bioneer catalog 2006 - Bioneer Catalog 2006.pdf · Enzymes Enzymes 98 Fig 2. Extension time was...
Transcript of bioneer catalog 2006 - Bioneer Catalog 2006.pdf · Enzymes Enzymes 98 Fig 2. Extension time was...
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6. Enzymes
Polymerases & Modifying Enzymes
Restriction Enzymes
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Polymerases & Modifying Enzymes
Taq DNA Polymerase
Product Size Contents Cat No.
Description: The Polymerase gene from Thermus aquaticus
(strain YT1) is cloned and expressed in E. coli, then Taq DNA
polymerase is produced with high purity. It is used in the
amplification and sequence testing of DNA through PCR. The
quality of the Taq DNA polymerase has been tested by the Activity
Test, SDS-Page, Endonuclease Test, etc.
Supplied with Enzyme
10 x Reaction buffer: 100 mM Tris-HCl, 400 mM KCl, 15 mM
MgCl2, pH 9.0.
Dilution Buffer: 20 mM Tris-HCl, 100 mM KCl, 0.5 mM EDTA,
1 mM DTT, 0.5 % Tween 20, 0.5 % IGEPAL CA-630 (pH 8.0.)
Storage Condition
20 mM Tris-HCl, 100 mM KCl, 0.5 mM EDTA, 1 mM DTT, 0.5 %
Tween 20, 0.5 % IGEPAL CA-630 (pH 8.0.) Store at -20
Concentration: 5 units/
Unit Definition
One unit is defined as the amount of enzyme required to catalyze
the incorporation of 10 nmole of dNTPs into an acid-insoluble form
in 30 minutes at 72
Activity Test
Working Range Test & Sensitivity Test
Using 1 unit of Taq DNA polymerase, the activity of the polymerase
was tested on human, bacterial, and lambda genomic DNA as
template. Each template DNA was serially diluted by tenfolds, with
different ranges.
A: Lambda genomic DNA (100 ng ~10 fg)
B: Bacteria genomic DNA (100 ng ~10 fg)
C: Human genomic DNA (100 ng ~10 pg)
Fig 1. Test of working range & sensitivity of Taq DNA
polymerase for each template. The PCR mixture was mixed with
serially diluted template DNA and 20 pmole of Forward/Reverse
primers, dNTPs Mix, and 10 x reaction buffer; diluted Taq DNA
polymerase was added just before PCR was carried out. From 10
fg to 100 ng of lambda genomic DNA was amplified, 10 fg to 100
ng of bacterial genomic DNA was amplified, and 10 pg to 100 ng of
human genomic DNA was amplified.
Lane 1: 100 ng of template DNA Lane 2: 10 ng of template DNA
Lane 3: 1 ng of template DNA Lane 4: 100 ng of template DNA
Lane 5: 10 ng of template DNA Lane 6: 1 ng of template DNA
Lane 7: 100 fg of template DNA Lane 8: 10 fg of template DNA
Examination of Taq DNA Polymerase working speed
The working speed of Taq DNA Polymerase wae tested by limiting
the extension time to 60 seconds. While conventional Taq DNA
polymerases had an average speed of ~ 60 nts/sec.
Enzyme Optimum
temp.( )
Half life(min, in
95 )
MgCl2(mM)
KCl(mM)
Optimum
pH (25 )
Taq 72 80 1.5 40 8.3
1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 1 2 3 4 5
A B C
Taq DNA Polymerase 250 units 10 mM dNTPs, 10 x reaction buffer with MgCl2 E-2010
Taq DNA Polymerase 500 units 10 mM dNTPs, 10 x reaction buffer with MgCl2 E-2011
Taq DNA Polymerase 1000 units 10 mM dNTPs, 10 x reaction buffer with MgCl2 E-2012
Taq DNA Polymerase 250 units 10 mM dNTPs, 10 x reaction buffer free MgCl2 , 20 mM MgCl2 E-2010-1
Taq DNA Polymerase 500 units 10 mM dNTPs, 10 x reaction buffer free MgCl2, 20 mM MgCl2 E-2011-1
Taq DNA Polymerase 1000 units 10 mM dNTPs, 10 x reaction buffer free MgCl2, 20 mM MgCl2 E-2012-1
Taq DNA Polymerase 250 units 10 x reaction buffer with MgCl2 E-2010-2
Taq DNA Polymerase 500 units 10 x reaction buffer with MgCl2 E-2011-2
Taq DNA Polymerase 1000 units 10 x reaction buffer with MgCl2 E-2012-2
Taq DNA Polymerase 250 units 10 x reaction buffer free MgCl2, 20 mM MgCl2 E-2010-3
Taq DNA Polymerase 500 units 10 x reaction buffer free MgCl2, 20 mM MgCl2 E-2011-3
Taq DNA Polymerase 1000 units 10 x reaction buffer free MgCl2, 20 mM MgCl2 E-2012-3
dNTP 1 ml 10 mM ( each 2.5 mM ) D-3001
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Fig 2. Extension time was limited to 60 sec and 1 ~ 10 kb of
lambda DNA target gene was amplified. Bioneer s Taq DNA
Polymerase amplified up to 10 kb of the target gene within the
60 sec extension period.
Lane M: 1 Kb DNA Ladder (D-1040, Bioneer)
Lane 1: 1 kb PCR Product Lane 2: 2 kb PCR Product
Lane 3: 3 kb PCR Product Lane 4: 4 kb PCR Product
Lane 5: 5 kb PCR Product Lane 6: 6 kb PCR Product
Lane 7: 7 kb PCR Product Lane 8: 8 kb PCR Product
Lane 9: 9 kb PCR Product Lane 10: 10 kb PCR Product
Efficiency test of Taq DNA polymerase
The amplification efficiency was examined according to the number
of cycles. PCR product after 30, 40, 50, 60 cycles was quantified
by fluorometer (Phamacia corp.) and the efficiency was determined
by the equation below 1 ng of human genomic DNA used.
Fig 3. M: 100 bp DNA Ladder (D-1030, Bioneer)
Lane 1: 30 cycle, Lane 2: 40 cycle, Lane 3: 50 cycle, Lane 4: 60
cycle
Nf= No (1+Y)n
- Nf: product copies following n cycles
- No: no. of template copies
- Y: efficiency of Taq DNA polymerase
- n: no. of PCR cycles
Reliability Test
To test the reproducibility of the produced batch, the Taq DNA
polymerase from each lot was serially diluted and the range of
amplification tested
Fig 4. Reliability Test PCR was carried out with Taq DNA
polymerase serially diluted to 1/5, 1/10, 1/15, 1/20 on lambda (A)
and human (B) genomic DNA as template. DNA carried out. The
amplification range of each lot is equal.
Lane 1: 5 U of Taq DNA pol. Lane 2: 1 U of Taq DNA pol.
Lane 3: 0.5 U of Taq DNA pol. Lane 4: 0.33 U of Taq DNA pol.
Lane 5: 0.25 U of Taq DNA pol.
Related Product
AccuPower TM PCR PreMix
AccuPower TM HL PCR PreMix
AccuPower TM RT PreMix
AccuPower TM RT/PCR PreMix
* This product is sold under licensing arrangements with Roche Molecular Systems, F.
Hoffmann-La Roche Ltd ( Roche ) and Perkin-Elmer Corporation. Purchase of this
product is accompanied by a license to use it in the Polymerase Chain Reaction
(PCR) process in conjunction with an Authorized Thermal Cycler.
<A> <B>
M 1 2 3 4
1 2 3 4 5 1 2 3 4 5
2.50E+12
2.00E+12
1.50E+12
1.00E+12
5.00E+11
0.00E+0020 30 40 50 60 70
Cycle No.
No. of Cycles Copy No. Efficiency of Taq DNA Pol.
30 6.20 E + 11 104%
40 1.55 E + 12 75%
50 1.99 E + 12 57%
60 2.08 E + 12 46%
M 1 2 3 4 5 6 7 8 9 10
Co
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o.
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Product Size Contents Cat No.
Description: HL Taq DNA Polymerase is themostable DNA
polymerase for high fidelity, long and high yield PCR. Also, it has
heat-stability, convenience, simplicity and reproducibility.
Feature & Benefits
HL Taq DNA polymerase is a unique enzyme containing
mixture of Taq DNA polymerase and the thermostable DNA
polymerase with proofreading activity. It suits well not only for
conventional PCR but also for the amplification of large DNA
template. The HL Taq DNA polymerase allows amplification of
DNA fragment up to 20 kb from lambda DNA.
HL Taq DNA polymerase offers about three times the fidelity
(1.89x10-6) and eight times the yield of other Taq DNA
polymerase.
Supplied with Enzyme
10 x Reaction Buffer (1ml): 100 mM Tris-HCl, 400 mM KCl, 15
mM MgCl2, pH 9.0
Dilution Buffer (1ml): 20 mM Tris-HCl, 0.5 mM EDTA, 1 mM
DTT,100 mM KCl, Stablizers, 50 % Glycerol, pH 8.0
dNTPs mixture (0.5 ml): 10 mM, each dNTP 2.5 mM
Storage Condition
20 mM Tris-HCl, 0.5 mM EDTA, 1 mM DTT, 100 mM KCl,
Stablizers, 50 % Glycerol pH 8.0, store at -20
Concentration: 5 units/
Unit Definition
One unit is defined at the amount of enzyme that will incorporates 10
nmol of dNTPs into acid-insoluble material in 30 minutes at 72 .
Quality Assurance
Nuclease activity is not detected after incubation of 1 ug of
substrate DNA-supercoiled plasmid and lambda/Hind III DNA - with
5 units of HL Taq DNA Polymerase in 50 uL reaction volume with
the supplied reaction buffer for 18 hrs at 37 and 70 .
General Reaction Condition [ 20 reaction volume ]
Reaction mixture
Template variable
Primer (forward) 5 ~ 10 pmoles
Primer (reverse) 5 ~ 10 pmoles
10 x Reaction buffer 2
10mM dNTPs mix. variable
Taq DNA Polymerase 0.5 ~ 1.0 unit
D.W variable
---------------------------------------------------------
Total 20
Note
In extremely long PCR, we recommend designing primers 35 ~
40 length bases containing 40 ~ 60% G+C residues with Tm of
68 ~ 72 and performing two step PCR - 94 for 30 sec, 68
for 1 min / 1 kb, 30 rounds.
Conventional
PCR (< 10kb)
DNA : 1 ~ 20 ngTemplate 5 ~ 50 ng Bacteria : 20 ~ 100 ng
Human : 100 ~ 500 ng
Primer 10 ~ 30 pmoles 5 ~ 20 pmoles
Extremely long PCR
(10 kb ~ 20 kb)
HL Taq DNA Polymerase
HLTaq DNA Polymerase 250 Units Each E-2018
HLTaq DNA Polymerase 1,000 Units Each E-2019
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HotStart Taq DNA Polymerase
Product Size Contents Cat No.
Description
HotStart Taq DNA Polymerase is a modified form of the recombinant
Taq DNA polymerase. The activity of HotStart Taq DNA polymerase
is completely inhibited when the temperature is below 70 . The
enzyme requires an activation step at 95 for 10~15 minutes . The
thermal activation prevents the extension of non-specifically
annealed primers and primer-dimers formed at low temperature
during PCR setup. The reaction condition of HotStart Taq DNA
polymerase can also be used without any modification for any PCR
reaction condition and cycling parameter of Taq DNA polymerase.
Activation step
HotStart Taq DNA polymerase needs to be activated for 15
minutes at 95 as initial step of PCR
Features and BenefitsHigh Specificity
High Sensitivity
Improve product yields
Reduce nonspecific background
Easy handling (Reliable Room-temperature setup)
ApplicationHotstart PCR
Low copy numbers PCR
Multiplex PCR
Quantitative PCR
Unit Definition
One unit is defined as the amount of enzyme required to catalyze
the incorporation of 10nmoles of dNTP into acid-insoluble material
in 30 minutes at 72 .
Quality Assurance
Nuclease activity is not detected after incubation of 1 ug of substrate
DNA - supercoiled plasmid and lambda/Hind III DNA - with 5 units
of HotStart Taq DNA Polymerase in 50 uL reaction volume with the
supplied Reaction buffer for 18 hr at 37 oC and 70 .
Supplied with10X Reaction Buffer : Contains Tris-HCl, KCl, 15 mM MgCl2, pH
9.0
dNTPs mixture : 10 mM, The concentration of each dNTPs is 2.5
mM.
Dilution and storage buffer : 20 mM Tris-HCl, 0.5 mM EDTA, 1
mM DTT, 100 mM KCl, Stabilizers, 50 % Glycerol, pH 8.0
Storage condition
HotStart Taq DNA polymerase, including buffers and reagents,
should be stored immediately upon receipt at -20 oC in a constant
temperature freezer.
Activity Test
Sensitivity test
The result of sensitivity comparison between One of the well-
known competitive companies (Q)’s HotStart and Bioneer’s
HotStart Taq DNA polymerase using Lambda and Human
Genomic DNA as template is shown below. 1 U of each enzymes
were used per reaction and each template DNA is continuously
diluted 10 times.
HotStart Taq DNA polymerase Competitive (Q) Hotstart Product
Fig 1. Sensitivity test Compared with Bioneer’s HotStart Taq
DNA polymerase and Qiagen’s Hotstart product. A) Using
Lambda DNA as a template, they were amplified up to 1kb
fragment length by having 5X10^8 copies to 5 copies B) Using
Human DNA as a template, they were amplified P53 gene of 400
bp fragment by having 5X10^4 copies to 50 copies. Lane 6 is
template negative.
Specificity test
Using Human genomic DNA (10ng) as a template, specificity
comparison is done between Taq DNA polymerase, HotStart Taq
DNA polymerase and the well-known Competitor company (Q)°Øs
HotStart and is confirmed specificity by amplifying P53 gene up to
1560bp (from 140bp to 1560) by using primer pairs that is over
65% GC content and 10?C Tm differences between each forward
and reverse primer.
HotStartTaq DNA Polymerase 250 Units 10mM dNTPs, 10 reaction buffer with MgCl2 E-2017
HotStartTaq DNA Polymerase 1000 Units 10mM dNTPs, 10 reaction buffer with MgCl2 E-2017-1
M 1 2 3 4 5 6 7 8 9 10
M 1 2 3 4 5 6 M 1 2 3 4 5 6
A)
B)
M 1 2 3 4 5 6 7 8 9 10
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Taq pol. HotStart Taq pol. Competitor CO.(Q)HotStart
Fig 2. Specificity Comparison between Taq, HotStart Taq DNA
Polymerase and Competitor company (Q)’s HotStart.
Enzyme dilution test
Amplify Human P53 of 600bp fragment after having HotStart Taq
DNA polymerase repeatedly diluted in the same sequential units
from 2U up to 0.125U(2U:1:0.5:0:25:0.125).
Fig 3: The amplification result of P53 gene (600bp) from Human
genomic DNA per units of HotStart Taq DNA polymerase.
M: 100bp DNA Ladder
Line 1: Enzyme 2 U Line 2: Enzyme 1 U
Line 3: Enzyme 0.5 U Line 4: Enzyme 0.25 U
Line 5: Enzyme 0.125 U Line 6: Enzyme negative
Multiplex PCR
1) Single & multiplex PCR using one template and five primer set.
Fig 4 : Result of Single & Multiplex PCR. Amplified P53 Gene from
Human Genomic DNA. The result of specific primer set is used in
each different lane from 1 to 5 and PCR reaction in one tube of the
primer set mixtures of lane from 1 to 5 in lane 7.
M : 100bp DNA Ladder
Line 1: 119bp fragment Line 2: 246bp fragment
Line 3: 314bp fragment Line 4: 445bp fragment
Line 5: 600bp fragment Line 6: Template DNA negative
Line 7: Multiplex PCR with primer mixtures from 1 to lane 5
2) Proceeded PCR reaction from one tube using 6 sets of primer
and 6 different types multiplex PCR of genomic DNA from six
different primer-template system.
HotStart Taq pol. HotStart Product (Company Q)
Fig 5 : Result of Single & Multiplex PCR.. From lane 1 to 6 is the
result of PCR reaction of genomic DNA and specific primer. Lane 7
is the PCR reaction in one tube of the primer set mixtures of lane
from 1 to 6. Lane 8 is the Competitor company(Q)’s multiplex PCR
kit..
M: 100bp DNA Ladder
Line 1: 750bp fragment Line 2: 590bp fragment
Line 3: 450bp fragment Line 4: 360bp fragment
Line 5: 260bp fragment Line 6: 150bp fragment
Line 7: Multiplex PCR with template DNA & primer mixtures from 1
to lane 6
Quantitative PCR
Real Time PCR Comparison data chart Using HotStart Taq DNA
polymerase and Company (Q)’s HotStart product. The result of
DNA Titration From 5 copies to 5x10^8 copies are as follows:
efficiency=0.30, Linearity = 0.999
HotStart Taq pol. HotStart Product (Company Q)
Fig 7. Quantitative PCR comparison between HotStart Taq-pol
and Company (Q)’s HotStart
Ordering Information
CAT.#
E-2017
E-2017-1
PRODUCT NAME
HotStart Taq DNA polymerase, 250Unit, 10 mM
dNTPs, 10 X reaction buffer with MgCl2
HotStart Taq DNA polymerase, 1000Unit, 10 mM
dNTPs, 10 X reaction buffer with MgCl2
M 1 2 3 4 5 6 7
M 1 2 3 4 5 6
M 1 2 3 4 5 6 7
M 1 2 3 4 5 6 7 M 1 2 3 4 5 6 7 1 2 3 4 5 6 7 M 1 2 3 4 5 6 7 M
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Pfu DNA Polymerase
Product Size Contents Cat No.
Description : Pfu DNA polymerase is a thermostable DNA
polymerase isolated from Pyrococcus furiosus Vc1. It catalyzes the
DNA-dependent polymerization of nucleotides into duplex DNA in
the 5 3 direction and exhibits 3 5 exonuclease (proof
reading) activity. Pfu DNA polymerase is the ideal choice for a
variety of techniques requiring high-fidelity DNA synthesis by PCR
reaction. It can apply to cloning, gene expression, site-directed
mutagenesis and etc.
Features and Benefits
High-Fidelity: 3 - 5 exonuclease(proofreading) activity
Thermostability: retaining 94-99% of its thermostable
activity after 1 hour at 95
Modified Nucleotide incorporation: incorporates the following
modified nucleotides: 7-deaza-dGTP, a-thionucleotide,
fluoresceinated and biotinylated nucleotides.
Terminal Transferase Activity: devoid of terminal transferase
activity and generates blunt-ended PCR products.
Applications
- Polymerase Chain Reaction (PCR)
- Primer Extension
- Gene Cloning
- Gene Expression
- Site-directed Mutagenesis
Supplied with Enzyme
10 x Reaction Buffer (1 m ): 200 mM Tris-HCl, 100 mM KCl,
100 mM (NH4)2SO4, 20 mM MgSO4, 1% Triton x -100, 1 mg/ml
Acetylated BSA, pH 8.8
Storage Condition
50 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, Stablizers, 50%
Glycerol, pH 8.2, store at -20
Concentration : 2.5 units/
Unit Definition
One unit is defined at the amount of enzyme that will incorporate 10
nmol of dNTP into acid-insoluble material in 30 minutes at 72
Quality Assurance
Nuclease activity is not detected after incubation of 1 ug of
substrate DNA-supercoiled plasmid and lambda/Hind III DNA- with
5 units of Pfu DNA Polymerase in 50 uL reaction volume with the
supplied 10 reaction buffer for 18 hrs at 37 and 70 .
Activity Test
Amplication of limiting amounts of template DNA: comparison of
Lambda DNA (A), Bacterial genomic DNA (B), Human genomic
DNA (C).
* Lambda DNA was amplified using 2.5 units of enzyme in 50 uL reaction volume.
M: 1 Kb DNA Ladder (D-1040, Bioneer)
Lane 1: 10 ng of Lambda DNA
Lane 2: 1 ng of Lambda DNA
Lane 3: 100 pg of Lambda DNA
Lane 4: 10 pg of Lambda DNA
Lane 5: 1 pg of Lambda DNA
Lane 6: 100 fg of Lambda DNA
Lane 7: 10 fg of Lambda DNA
* Bactierial DNA was amplified using 2.5 units of enzyme in 50 uL reaction volume.
M: 1 Kb DNA Ladder (D-1040, Bioneer)
Lane 1: 10 ng of Bacterial DNA
Lane 2: 1 ng of Bacterial DNA
Lane 3: 100 pg of Bacterial DNA
Lane 4: 10 pg of Bacterial DNA
Lane 5: 1 pg of Bacterial DNA
Lane 6: 100 fg of Bacterial DNA
* Human DNA was amplified using 2.5 units of enzyme in 50 uL reaction volume.
M: 1 Kb DNA Ladder (D-1040, Bioneer)
Lane 1: 10 ng of Human DNA
Lane 2: 1 ng of Human DNA
Lane 3: 100 pg of Human DNA
Lane 4: 10 pg of Human DNA
Lane 5: 1 pg of Human DNA
Lane 6: 100 fg of Bacterial DNA
Amplification of fragments ranging from 500 bp to 5 kb from
template Lambda DNA 100 pg with 2.5 units of Pfu DNA
Polymerase.
M: 1 Kb DNA Ladder
(D-1040, Bioneer)
Lane 1: amplification of 500 bp
fragment
Lane 2: amplification of 1.0 Kb
fragment
Lane 3: amplification of 2.0 Kb
fragment
Lane 4: amplification of 3.5 kb fragment
Lane 5: amplification of 5.0 kb fragment
M 1 2 3 4 5 6 7
M 1 2 3 4 5 6
M 1 2 3 4 5
A
B
C
Pfu DNA Polymerase 250 Units 10 x reaction buffer, without dNTPs E-2015
Pfu DNA Polymerase 1,000 Units 10 x reaction buffer, without dNTPs E-2016
dNTP 1 ml 10 mM ( each dNTPs, 2.5 mM ) D-3001
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M-MLV Reverse Transcriptase
Product Size Contents Cat No.
T7 RNA Polymerase
Description : Moloney Murine Leukemia Virus (M-MLV) Reverse
Transcriptase is an RNA-dependent DNA polymerase. This
enzyme is able to use an RNA molecule as a template and
synthesize a double-strand DNA .
Source
M-MLV Reverse Transcriptase is isolated from a E.coli strain
containing a recombinant clone.
Applications
First-strand synthesis of cDNA from RNA molecules.
Supplied with Enzyme
5 x Reaction Buffer (0.5 ml): 250 mM Tris-HCl, 150 mM KCl,
40 mM MgCl2, pH 8.3
100 mM DTT: 0.3 ml
Storage Condition
20 mM Tris-Cl (pH7.6), 150 mM NaCl, 0.1 mM EDTA, 1 mM DTT,
0.1% IGEPAL CA-630, 50% Glycerol, store at -20
Concentration: 200 units/
Unit Definition
One unit is defined as the amount of enzyme required to
incorporate 1 nmole of dTTP into acid-precipitable material in 10
min at 37 using oligo d (T) as template primer.
Quality Assurance
DNase and RNase activity is not detected after incubation of 1ug of
DNA and RNA with 200 units of M-MLV Reverse Transcriptase for
3 hours at 37 ~ 42 .
References
1. Gerard, G. F. et al. (1975) J. Virol. 15, 785 - 797
2. Roth, M. et al. (1985) J. Biol. Chem. 260, 9326 - 9335
3. Sambrook, J. Fritsch, E. F. and Maniatis, T. (1989) Molecular
Cloning: A Laboratory Manual, second edition, Cold Spring
Harbor, New York, 8. 64.
Related Products
AccuPower TM RT PreMix
AccuPower TM RT/PCR PreMix
AccuRapid TM PCR cDNA Library Kit
Product Size Contents Cat No.
Description: T7 RNA Polymerase is a DNA dependent RNA
polymerase with a highly specificity for the initiation of transcription
at T7 RNA polymerase promoters. It is widely used for the rapid
synthesis in vitro of specific RNAs.
Source
T7 RNA Polymerase is isolated from E.coli cells containing the
ligase gene cloned from T7 bacteriophage.
Applications
Synthesis of RNA transcripts for northern hybridization and
southern hybridization probes.
RNA generation for studies of RNA structure, procession and
catalysis
Supplied with Enzyme
10 x Reaction Buffer (1 ml): 400 mM Tris-Cl (pH 8.0), 60 mM
MgCl2, 20 mM Spermidine
100 mM DTT (0.5 ml)
DEPC-DW (1ml)
Storage Condition
200 mM Na-phosphate (pH 7.7), 10 mM NaCl, 1 mM EDTA, 1 mM
DTT, 0.02 % Triton x -100, 0.08% Tween-20, 50% Glycerol, Store
at -20
Concentration: 0.1 units/
Unit Definition
One unit of enzyme catalyzed incoporation of 1 nmoles of
[3H]ATPs into acid insoluble form in 60 min at 37
Quality Assurance
Nuclease Contamination Assay: No altered was detected after
incubation of 1 ug of substrate DNA with 500 units of T7 RNA
polymerase for 18 hrs in 37
Protease Contamination Assay: No altered pattern was
detected after in cubation of 2,000 units of T7 RNA polymerase
for 18 hrs in 37
M-MLV Reverse Transcriptase 10,000 Units 5 x reaction buffer, 0.5 ml, 100 mM DTT, 0.3 ml E-3121
M-MLV Reverse Transcriptase 50,000 Units 5 x reaction buffer, 2.5 ml, 100 mM DTT, 1.5 ml E-3122
T7 RNA Polymerase 2,000 Units 10 x reaction buffer, 1 ml, 100 mM DTT, 0.5 ml, DEPC-DW, 1 ml E-3041
T7 RNA Polymerase 10,000 Units 10 x reaction buffer, 5 ml, 100 mM DTT, 2.5 ml , DEPC-DW, 5 ml E-3042
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DNA Polymerase 1, Large (klenow) Fragment
Product Size Contents Cat No.
Description
DNA Polymerase I, Large (Klenow) Fragment is derived from E.coli
DNA polymerase that it has the function of Polymerization an 3’ 5’
exonuclease but it has deficient activity of 5’ 3’ exonuclease.
Source
DNA Polymerase I, Large (Klenow) Fragment is isolated from an E.coli
strain carrying the gene of the enzyme on a plasmid.
Applications
DNA sequencing by the dideoxy method
Random priming labeling
Second strand synthesis in oligonucleotide directed mutagenesis
Reaction Condition
10 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 1 mM DTT, Incubate at 37 .
Storage Conditions
20 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM EDTA, 10 mM 2-
mercaptoethanol, 50% Glycerol,
Store at -20
Unit Definition
One unit of enzyme is defined as the amount of enzyme required to
convert 10 nmoles of dNTPs to an acid-soluble form in 30 min at 37 .
Quality Assurance
Nuclease Contamination Assay: No altered was detected after
incubation of 1ug of substrate DNA with 100units of DNA
Polymerase I (Klenow Fragment) for 18 hrs in 37 .
Protease Contamination Assay: No altered pattern was detected
after incubation of 100 units of DNA polymerase I (Klenow Fragment)
for 18 hrs in 37 .
Concentration : 5 units/
DNA Polymerase I 200 U Each E-3021DNA Polymerase I 1,000 U Each E-3022
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Description: T4 DNA Ligase catalyzes the formation of a
phosphodiester bond between juxtaposed 5 phosphate and 3
hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt-
end and cohesive-end termini as well as repair single stranded
nicks in duplex DNA, RNA, or DNA/RNA hybrids.
Source: T4 DNA Ligase is isolated from a recombinant E.coli
strain.
Applications: Joining double-stranded DNA with cohesive or
blunt ends
Supplied with Enzyme
10 x Reaction Buffer (1 ml) : 500mM Tris-HCl (pH 7.8),100mM
MgCl2, 50mM DTT, 10mM ATP, 25 ug/ml BSA
Storage Condition
10mM Tris-HCl (pH 7.5), 50mM KCl, 1mM EDTA, 10 mM 2-
mercaptoethanol, 50% glycerol, store at -20 .
Concentration: 1 unit /
Unit Definition: 0.01 Weiss unit of enzyme is defined as the
amount of enzyme required to give 90% ligation of Hind lll
fragments of lambda DNA in 30 min at 16 in 20 of the assay
mixture.
Quality Assaurance: No altered was detected after incubation of
1 substrate DNA with 10 units of T4 DNA Ligase in 20 ul reaction
volume with the supplied reaction buffer for 18 hrs at 37
Note Store the buffer in small aliquots at -20 to minimize
degradation of the ATP and DTT
Ligation Test
Product Size Contents Cat No.
Lane 1: DNA fragment (digested with EcoR V)
Lane 2,3,4,5: T4 DNA Ligase 1 unit, 16 , 10 min, 20 min, 30 min, 60 min
Lane 6,7,8,9: T4 DNA Ligase 1 unit, 25 , 10 min, 20 min, 30 min, 60 min
Lane 10,11,12,13: T4 DNA Ligase 1 unit, 37 , 10 min, 20 min, 30 min, 60 min
Lane 14: lambda DNA (digested with Hind III)
Lane 15,16,17: T4 DNA Ligase 0.01 unit , 16 , 10 min, 20 min, 30 min
Lane 18,19,20: T4 DNA Ligase 0.01 unit , 25 , 10 min, 20 min, 30 min
Lane 21,22,23: T4 DNA Ligase 0.01 unit , 37 , 10 min, 20 min, 30 min
Blunt-end ligation Cohesive-end ligation
1 2 3 4 5 6 7 8 9 10 12 13 14 15 16 17 18 19 20 21 22 23 24
T4 DNA Ligase
T4 DNA Ligase 100 Weiss unit Each E-3061
T4 DNA Ligase 500 Weiss unit Each E-3062
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Thermostable Thermus Filiformis (Tfi) DNA Ligase
Product Size Contents Cat No.
Description : Tfi DNA Ligase plays role of formatting
phosphodiester bond by connecting double stranded DNA or
connecting oligonucleotides 3’-hydroxyl end and 5’-phosphate end
together. Especially the reaction temperatures are between 45
and 65 compared to DNA Ligases such as T4 DNA Ligse, E.coli
DNA Ligase, etc. It is the higher temperature that keeps safe and
active. In addition, Tfi DNA Ligase is possible to use in ligation
reaction in high reaction temperature requirement.
Source
Tfi DNA Ligase is isolated from E.coli cells containing the ligase
gene cloned from Thermus filiformis.
Applications
Ligase Chain Reaction (LCR)
Oligonucleotide Ligation Assay (OLA)
Mutagenesis by Incorporation of a phosphorylated oligo during
PCR Amplication
Simultaneous Mutagenesis of Multiple Sites
Supplied with Enzyme
10 x Reaction Buffer (1 ml): 300 mM Tris-HCl (pH8.3), 250 mM
KCl, 50 mM MgCl2, 5 mM NAD
1 x Dilution Buffer (1 ml): 10 mM Tris-HCl (pH7.6), 0.1 mM
EDTA, 50 mM KCl, 1 mM DTT, 200 ug/mL acetylated BSA,
50% Glycerol
Storage Condition
20 mM Tris-HCl (pH7.6), 2 mM MgCl2, 1 mM EDTA, 1 mM DTT,
0.5% Tween -20, 0.5% IGEPAL CA-630, 50% Glycerol, store at -
20 .
Concentration : 20 units/
Unit Definltion
One unit of Tfi DNA Ligase is defined as the amount of enzyme
required to give 50% ligation of the 12 base pair cohesive ends of 1
ug of PspEI digested lambda DNA in 10min at 45 .
Activity Assay Conditions
The activity assay is carried out in a 20 reaction containing 1 ug
of PspEI digested lambda DNA and 1 x Tfi DNA ligase reaction
buffer. After incubation at 45 for 10 min., the reaction is
terminated by addition of stop solution (40%(w/v) sucrose, 50 mM
EDTA and 0.25% bromophenol blue). Then heat at 70 for 10
min. and immediately load on a 0.8% agarose gel.
Stability
The half-life of the enzyme in 1 x reaction buffer is more than 1
hour at 95 and 55 hours at 65 .
Note : Tfi DNA Ligase should not be used as a substitute for other
DNA ligases, i.e., T4 DNA Ligase.
References
1. Barany, F. (1991) Proc. Natl. Acad. Sci. USA, 88, 189 - 193.
2. Landegren, U. et al.(1988) Science 241, 1077 - 1080
3. Michael, Scott F. (1994) Biotechniques 16:3, 410 - 412.
4.Gerard J. A. et al. (1993) Biotechniques 15:1, 172 - 178.
1 2 3 4 5 6
LigationProduct(1+4)
1. Fragment
4. Fragment
Lane 1: DNA/PspE I (control)
Lane 2: Incubate at 45 , 10 min
Lane 3: Incubate at 50 , 10 min
Lane 4: Incubate at 55 , 10 min
Lane 5: Incubate at 60 , 10 min
Lnne 6: Incubate at 65 , 10 min
Control Condition: Incubate the reaction containing ligase 1 unit and
1 ug DNA[lambda PspEI] at each temperature for 10 min.
Test of temperature for ligation (45 ~ 65 )
Tfi DNA Ligase 2,000 Units 10 x reaction buffer, 1 ml, 1 x dilution buffer, 1 ml E-3111
Tfi DNA Ligase 10,000 Units 10 x reaction buffer, 5 ml, 1 x dilution buffer, 5 ml E-3112
Enzymes
En
zyme
s
107
1 2 3 4 5 6 7 8 9 10
Lane 1: DNA/PspE I (control) Lane 2: Incubate at 95 , 10 min
Lane 3: Incubate at 95 , 20 min Lane 4: Incubate at 95 , 30 min
Lane 5: Incubate at 95 , 40 min Lane 6: Incubate at 95 , 50 min
Lane 7: Incubate at 95 , 60 min Lane 8: Incubate at 95 , 70 min
Lane 9: Incubate at 95 , 80 min Lane 9: Incubate at 95 , 90 min
Control condition : Incubate the enzyme at 95 each time. Andthen add 1unit ligase to a 20 reaction containing 1ugDNA[lambda PspEI] and incubate the mixture at 45 for 10 min.
1 2 3 4 5 6 7 8 9 10 11 12 13 14
Lane 1: DNA/PspE I (control) Lane 2: Incubate at 65 , 6 hrs
Lane 3: Incubate at 65 , 12 hrs Lane 4: Incubate at 65 , 18 hrs
Lane 5: Incubate at 65 , 24 hrs Lane 6: Incubate at 65 , 30 hrs
Lane 7: Incubate at 65 , 36 hrs Lane 8: Incubate at 65 , 42 hrs
Lane 9: Incubate at 65 , 48 hrs Lane 10: Incubate at 65 , 54 hrs
Lane 11: Incubate at 65 , 60 hrs Lane 12: Incubate at 65 , 66 hrs
Lane 13: Incubate at 65 , 72 hrs Lane 14: Incubate at 65 , 78 hrs
Control condition: Incubate the enzyme for each time at 65
and then add 1 unit ligase to a 20 reaction containing
1 ug DNA[lambda PspEI] and incubate the mixture at 45 for 10
min.
Heat stability test at 95 and 65
Enzymes
En
zym
es
108
Description
Tte Inorganic Pyrophosphatase (Ppase) is enzyme to make
orthophosphate form by hydrolyzing Inorganic Pyrophosphatase.:
P2O7-4+H2O -> 2HPO4
-2
Source
Tte Inorganic Pyrophosphatase (PPase) is isolated from Thermus
Thermophilus B35.
Storage Buffer
20 mM Tris-HCl (pH 8.0), 0.2 % Triton X-20, 100 mM KCl, 0.1 mM
EDTA,
50 % Glycerol, store at -20
Unit Definition
One unit is the amount of enzyme that will generate 40nmoles of
phosphate per minute from pyrophosphate under standard
reaction conditions (50 mM Tris-HCl (pH8.5), 1 mM MgCl2, 0.32
mM PPi) at 65 .
Concentration
300 units/ml
Reference : Heinonen, J. K, and Lahti, R. J. (1981) Analytical
Biochemistry 113, 313-317
Product Unit Contents Cat No.
PPase 0.5 Each E-3071
PPase 2.5 Each E-3072
Tte Inorganic pyrophophatase(PPase)
BstSE(Nickase) GAGTC (4/-)
Description
BstSE is site specific endonuclease that inltiates cleavage only in
single strand of double strand DNA
Source
BstSE is isolated from Bacillus Stearothermophilus SE-589.
Reaction Buffer
10 mM Tris-HCl (pH 8.5), 10 mM MgCl2, 150 mM KCl, 1 mM DTT,
Incubate at 55 .
Storage Conditions
10 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM EDTA, 10 mM 2-
mercaptoethanol, 50 % Glycerol, store at -20
Ligation and Recutting
After 5-fold overdigestion with BstSE, 90 % of the DNA fragments
can be ligated and recut.
Concentration
5 units/
Note : BstSE catalyzes a single-strand nick on the 3 side of the
recognition sequnece.
Product Unit Contents Cat No.
BstSE (Nickase) GAGTC (4/-) 50 Each E-2181
BstSE (Nickase) GAGTC (4/-) 250 Each E-2182
Enzymes
En
zyme
s
109
Bovine Serum Albumin(BSA), Acetylated
Description
Acetylated BSA is an enzyme stabilizer that is essential carrier
protein to remove contaminants such as nuclease and protease.
Acetylation process does not affect specific character of BSA nor
PCR but it removes nuclease’s activation very accurately. Add
BSA to be 5~200 /ml as a final concentration.
Source Bos Taurus
Storage Conditions
10 mM Tris-HCl (pH 7.5), Store at -20
Concentration
20 mg/ml
Reference
Gonzales, N., Wiggs, J., and Chamberlin, M. J. (1977) Arch.
Biochem. Biophys. 182, 404 - 408
Product Unit Contents Cat No.
BSA 20 mg Each E-3091
BSA 100 mg Each E-3092
Enzymes
En
zym
es
110
AccuRapidTM DNA Ligase
Description
AccuRapidTM DNA Ligase is a product can ligate cohesive-ended
DNA fragment and blunt-ended DNA fragment within 5 minutes
that reaction optimal temperature is from 15 to 25 .
AccuRapidTM DNA Ligase Buffer is the buffer type of product that
can give high efficient ligation result through cohesive-end and
blunt-end DNA fragment reaction. Ligation product is possible to
use directly into transformation.
Cat No. K-7101 Contents: For 50 ligations
50 unit(1 unit/ )
AccuRapidTM DNA Ligase(from T4 bacteriophage)
0.5 ml
2x AccuRapidTM DNA ligation buffer
Cat No. K-7102 Contents : For 150 ligations
50 unit x 3 (1 unit/ )
AccuRapidTM DNA Ligase(from T4 bacteriophage)
0.5 ml x 3
2x AccuRapidTM DNA ligation buffer
Storage Condition : Store at -20
Quality Control Assay: The kit is pre-tested in a ligation
according to the standard protocol; 500 ng of blunt-ended 500 bp
insert DNA was ligated to 100 ng of pUC19 DNA digested with
Sma I. Vector DNA fragments should most preferably be
dephosphorylated to minimize self-circularization. Yield of white
colonies after transformation is > 90%
Quality Assaurance: Nuclease activity is not detected after
incubation of 1 of substrate DNA with 10 units of AccuRapid
DNA Ligase in 20 reaction volume with the supplied Reaction
buffer for 18 hrs at 37
Reference
feiffer, B. H. and Zimmerman, S,B., Nucleic Acids Research, 11,
7853 ~ 7871, 1983.
Hayashi., Nakazawa, M., Ishizaki, Y., Hiraoka, N., and Obayashi,
A., Nucleic Acids Research, 13, 7979 ~ 7992, 1985.
Ligation test of reaction temperatures
Lane 1: 1 Kb DNA Ladder(D-1040)
Lane 2: EcoR V digested DNA fragments
Lane 3: Ligation reaction with T4 DNA Ligase reaction
buffer(E-3061) at 37 for 5 min
Lane 4: Ligation reaction with AccuRapid DNA Ligation
Kit at 16 for 5 min
Lane 5: Ligation reaction with AccuRapid DNA Ligation
Kit at 25 for 5 min
Lane 6: Ligation reaction with AccuRapid DNA Ligation
Kit at 37 for 5 min
Product Size Cat No.
AccuRapidTM DNA Ligation Kit 50 ligations K-7101
AccuRapidTM DNA Ligation Kit 150 ligations K-7102
Enzymes
En
zyme
s
111
B
B
I
B
O
I
I
N
I
B
I
N
B
I
N
R
B
I
R
N
O
R
B
R
R
O
O
B
I
O
O
N
R
O
R
Bsp13 I
B
O
I
B
B
I
O
I
B
B
B
O
I
I
I
I
Acc16 I
Acc65 I
Acc113 I
AccB1 I
AccB7 I
AccBS I
Acl I
AclN I
AclWI
Acs I
Afe I
Alu I
Ama87 I
Apa I
AsiA I
AspLE I
AspS9 I
AsuC2 I
AsuHP I
AsuNH I
BamH I
Bbv12 I
Bgl I
Bgl II
Bme18 I
Bpu14 I
Bsa29 I
Bsc4 I
Bse1 I
Bse3D I
Bse8 I
Bse21 I
Bse118 I
BseP I
BseX3 I
Bsp13 I
Bsp19 I
Bsp1720 I
BspA2 I
BssNA I
BssT1 I
Bst2B I
Bst2U I
Bst4C I
BstAC I
BstAP I
BstBA I
BstDE I
BstDS I
BstF5 I
BstH2 I
BstHP I
100
100
0-10
100
50-75
25-50
0-10
25-50
0-10
100
75-100
50-75
100
0-10
25-50
50-75
100
125-50
75-100
0-10
75-100
75-100
100
25-50
75-100
25-50
50-75
100
0-25
50-75
75-100
0-10
75-100
50-75
50-75
50-75
100
50-75
50-75
100
100
25-50
50-75
25-50
100
100
100
50-75
25-50
50-75
10-25
25-50
75-100
10-25
100
50-75
50-75
100
100
75-100
100
0-25
100
75-100
0-10
100
25-50
25-50
50-75
100
25-50
75-100
25-50
10-25
25-50
10-25
0-25
75-100
75-100
25-50
100
75-100
50-75
50-75
25-50
50-75
10-25
0-10
10-25
75-100
100
75-100
10-25
100
10-25
100
75-100
25-50
25-50
10-25
100
100
100
100
75-100
25-50
25-50
10-25
100
75-100
0-10
75-100
50-75
50-75
25-50
75-100
50-75
25-50
75-100
75-100
50-75
50-75
50-75
50-75
100
10-25
50-75
10-25
25-50
100
100
75-100
75-100
100
100
75-100
50-75
100
25-50
50-75
25-50
100
75-100
50-75
25-50
25-50
100
75-100
75-100
25-50
25-50
100
75-100
50-75
50-75
50-75
50-75
10-25
50-75
50-75
10-25
75-100
0-10
100
75-100
25-50
0-25
100
0-25
50-75
100
0-25
50-75
75-100
10-25
100
25-50
0-10
50-75
0-10
0-25
50-75
25-50
75-100
75-100
10-25
25-50
100
0-10
50-75
10-25
25-50
0-10
50-75
75-100
50-75
10-25
75-100
75-100
75-100
75-100
25-50
25-50
75-100
0-10
75-100
50-75
0-10
25-50
75-100
0-10
10-25
25-50
25-50
0-10
10-25
0-10
50-75
75-100
0-25
75-100
0-10
50-75
100
75-100
10-25
100
0-10
75-100
100
0-10
100
100
25-50
50-75
50-75
25-50
25-50
75-100
0-10
100
75-100
100
75-100
75-100
50-75
25-50
75-100
25-50
10-25
50-75
10-25
50-75
75-100
75-100
25-50
50-75
25-50
0-10
10-25
37
37
37
37
37
37
37
37
37
50
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
55
65
60
60
37
65
50
50
50
37
37
37
37
60
60
60
65
37
60
65
60
65
65
65
37
’
EnzymeOptimalBuffer B I O N R
OptimalTemp.
Activity in AccuCut buffers (%)
Restriction Enzymes
Enzymes
En
zym
es
112
N
N
R
N
O
I
O
I
O
I
EcoR I
B
N
B
N
I
I
I
R
O
B
R
N
I
N
O
O
R
N
N
B
N
R
B
N
I
N
O
N
I
I
I
I
R
O
N
R
I
R
O
N
N
I
R
O
N
I
B
I
O
R
B
R
I
N
N BstSE
Bst MS I
BstNS I
BstSF I
BstSN I
BstX2 I
Bsu6 I
BsuR I
CciN I
Dra I
DseD I
EcoR I
EcoR V
Ege I
Erh I
Fau I
FauND I
Fok I
FriO I
Fsp4H I
Hae III
Hind III
Hinf I
Hpa II
HspA I
Kpn I
Ksp22 I
Kzo9 I
Mlu I
Mly113 I
MroN I
MroX I
Msp I
MspR9 I
Nru I
NruG I
Ple19 I
Pme55 I
Psp124B I
PspE I
PspL I
PspN4 I
PspOM I
PspPP I
Pst I
Pvu I I
Rsa I
Sal I
Sbf I
SfaN I
Sfi I
Sfr274 I
Sfr303 I
Sma I
Smi I
Sph I
Sse9 I
Ssp I
Tru9 I
Tth111 I
Vha464 I
Vne I
Vsp I
Xba I
Xma I
Zsp2 I
N BstSE
10-25
10-25
50-75
10-25
10-25
10-25
50-75
75-100
75-100
50-75
100
100
50-75
100
0-10
50-75
25-50
0-10
75-100
50-75
100
75-100
25-50
25-50
25-50
50-75
50-75
25-50
10-25
0-10
100
75-100
50-75
100
10-25
25-50
10-25
0-10
50-75
10-25
25-50
0-10
10-25
25-50
25-50
50-75
25-50
0-10
75-100
25-50
50-75
10-25
0-10
75-100
75-100
50-75
50-75
100
10-25
10-25
25-50
100
50-75
0-10
25-50
10-25
50-75
75-100
50-75
50-75
25-50
100
50-75
100
75-100
100
50-75
25-50
75-100
10-25
50-75
100
100
100
25-50
50-75
0-10
75-100
50-75
100
75-100
25-50
50-75
10-25
50-75
10-25
25-50
75-100
75-100
10-25
75-100
100
25-50
75-100
50-75
100
100
100
100
25-50
25-50
75-100
0-10
100
0-10
25-50
75-100
75-100
100
25-50
50-75
75-100
100
50-75
100
75-100
25-50
25-50
75-100
100
25-50
0-10
75-100
50-75
25-50
50-75
100
50-75
100
50-75
100
75-100
75-100
25-50
75-100
25-50
25-50
75-100
50-75
75-100
50-75
100
25-50
75-100
50-75
50-75
25-50
100
100
10-25
25-50
50-75
50-75
75-100
50-75
10-25
25-50
75-100
50-75
100
50-75
75-100
10-25
10-25
25-50
25-50
100
50-75
10-25
50-75
25-50
100
75-100
50-75
0-10
25-50
100
75-100
50-75
25-50
50-75
100
25-50
10-25
75-100
50-75
50-75
0-10
100
100
75-100
100
75-100
50-75
75-100
25-50
75-100
75-100
50-75
0-10
100
10-25
100
50-75
50-75
75-100
50-75
75-100
10-25
25-50
100
50-75
100
50-75
50-75
0-10
100
100
50-75
100
50-75
0-10
100
75-100
100
75-100
100
75-100
10-25
75-100
50-75
10-25
25-50
100
0-10
75-100
10-25
75-100
100
100
0-10
25-50
25-50
100
75-100
75-100
75-100
50-75
10-25
0-10
75-100
75-100
100
0-10
10-25
10-25
100
0-10
0-10
10-25
25-50
75-100
25-50
25-50
75-100
50-75
10-25
25-50
0-10
10-25
25-50
10-25
100
25-50
0-10
100
10-25
25-50
25-50
50-75
50-75
100
10-25
10-25
50-75
50-75
100
75-100
10-25
25-50
10-25
10-25
25-50
25-50
10-25
0-10
10-25
100
25-50
0-10
100
0-10
100
25-50
50-75
10-25
0-10
100
75-100
50-75
25-50
25-50
10-25
25-50
100
50-75
100
0
25-50
10-25
50
37
60
65
60
37
37
37
37
37
37
37
37
37
55
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
37
50
50
37
25
37
37
55
37
65
65
37
37
37
37
37
37
55
Enzymes
En
zyme
s
113
Restriction Enzymes Isoschizomer Contents
Acc16 I
Acc65 I
Acc113 I
AccB1 I
AccB7 I
AccBS I
Acl I
AclN I
AclW I
Acs I
Afe I
Alu I
Ama87 I
Apa I
AsiA I
AspLE I
AspS9 I
AsuC2 I
AsuHP I
AsuNH I
BamH I
Bbv12 I
Bgl I
Bgl II
Bme18 I
Bpu14 I
Bsa29 I
Bsc4 I
Bse1 I
Bse3D I
Bse8 I
Bse21 I
Bse118 I
BseP I
BseX3 I
Bsp13 I
Bsp19 I
Bsp1720 I
BspA2 I
BssNA I
BssT1 I
Bst2B I
Bst2U I
BstAC I
BstAP I
BstBA I
BstDE I
BstDS I
BstF5 I
BstH2 I
BstHP I
BstMC I
BstNS I
BstSF I
BstSN I
BstX2 I
Bsu6 I
BsuR I
Aos I, Avi II, Fsp I, Mst I, Nsb I
Asp718 I, Sth I
Dpa I, Eco255 I, Sca I
Ban I, Hgic I, BbvB I, Eco64 I, MspB4 I
Acp II, Asp10H II, Esp1396 I, Van91 I, PflM I
BsrB I, Mbi I
Psp1406 I
Spe I
Bin I, Alw I, BspP I
Apo I, Fsi I, Xap I
Ait I, Aor51H I, Fun I, Eco47 III
Mlt I
Aqu I, Ava I, Bco I, BsoB I, Eco27K I, Eco88 I, NspII, NspSA I
Ppe I
Age I, PinA I, BshT I
Cfo I, FnuD III, Hha I
Asu I, Avc I, BspB II, Bsu54 I, Cfr13 I, Sau96 I
Aha I, Bcn I, Cau II, HgiS22 I, Nci I
Hph I
Nhe I, PstNH I
AccEB I, Ali I, Bna I, Bst I, NspSA IV, Sur I
Alw21 I, AspH I, Bsh45 I, HgiA I, BsiHKA I
Tsp8E I
Ncr I, NspMAC I, Pae2k I, Pae18k I
Afl I, Ava II, Bme216 I, Cau I, Eco47 I, HgiB I, HgiE I, Sin I
Acp I, Asp10H I, Asu II, Bsp119 I, BstB I, Csp45 I, Fsp II, Lsp I, Nsp V, Sfu I, Ssp1 I
Aag I, Ban III, Bci29 I, Bsc I, Bsp106 I, Bsu15 I, Cla I
BsiY I, Bsl I, BsaL I
BseN I, Bsr I, BsrS I, Bst11 I, Tsp1 I
BsrD I, BseM I
BsaB I, Bsh1365 I, BsiB I, BsrBR I, Mam I
Aoc I, Axy I, Bst29 I, Bst30 I, Bsu36 I, Cvn I, Eco81 I, Mst II,Sau I, SshA I
Bco118, BsrF I, BssA I, Cfr10 I
BssH II, Pau I
Aaa I, BstZ I, Eag I, EclX I, Eco52 I, Xma III
Acc III, BseA I, BsiM I, BspE I, BspM II, Bsu23 I, Kpn2 I, Mro I, Pta I
Nco I
Blp I, Bpu1102 I, Cel II, Esp I
Avr II, AvrB II, Bln I
BspM90 I, BstBS I, Bst1107 I, Sna I, Xca I
EcoT14 I, Eco130 I, Erh I, ErhB9 II, Sty I
Bsi I, BssSI
Aor I, Apy I, BseB I, Bse16 I, Bse17 I, Bse24 I, BspN I, Bst2 I, BstN I, BstO I, EcoR II,Fsp1604 I, Mva I,
Sth117 I, Zan I
Acy I, Aha I, Asu III, Bbi II, BsaH I, Hgi I, Hin1 I, Msp17 I, Pam II
ApaB I
BsaA I, MspY I
Dde I
Dsa I
BseG I
AccB2 I, Bsp143 II, Hae II
Hpa I
BsaO I, Bsh1285 I, BsiE I, Mcr I
Nsp I, NspH I
BdisS I, LlaB I, Sfc I, Sfe I
Eco105 I, Sna B I
BstY I, Mfl I, Tru201 I, Xho II
Bco5 I, Bco116 I, BseZ I, Ear I, Ksp632 I
Bim19 II, Bsh I, BspK I, BspR I, Bsp211 I, Dsa II, FnuD I, Hae III, Pla I, Sbv I, Sfa I
TGC GCA
G GTACC
AGT ACT
G G(C/T)(G/A)CC
CCANNNN NTGG
GAG CGG
AA CGTT
A CTAGT
GGATCNNNN
(G/A) AATT(C/T)
AGC GCT
AG CT
C (C/T)CG(G/A)G
GGGCC C
A CCGGT
GCG C
G GNCC
CC (C/G)GG
GGTGANNNNNNNN
G CTAGC
G GATCC
G(A/T)GC(A/T) C
GCCNNNN NGGC
A GATCT
G G(A/T)CC
TT CGAA
AT CGAT
CCNNNNN NNGG
ACTG G
GCAATGNN
GATNN NNATC
CC TNAGG
(G/A) CCGG(C/T)
G CGCGC
C GGCCG
T CCGGA
C CATGG
GC TNAGC
C CTAGG
GTA TAC
C C(A/T)(A/T)GG
C TCGTG
CC (A/T)GG
G(G/A) CG(C/T)C
GCANNNN NTGC
(C/T)AC GT(G/A)
C TNAG
C C(A/G)(C/T)GG
GGATGNN
(G/A)GCGC (C/T)
GTT AAC
CG(G/A)(C/T) CG
(G/A)CATG (C/T)
C T(G/A)(C/T)AG
TAC GTA
(G/A) GATC(C/T)
CTCTTCN
GG CC
BioneerEnzyme
Isoschizomer
Enzymes
En
zym
es
114
CciN I
Dra I
DseD I
EcoR I
EcoR V
Ege I
Erh I
Fau I
FauND I
Fok I
FriO I
Fsp4H I
Hae III
Hind III
Hinf I
Hpa II
HspA I
Kpn I
Ksp22 I
Kzo9 I
Mlu I
Mly113 I
MroN I
MroX I
Msp I
MspR9 I
Nru I
NruG I
Ple19 I
Pme55 I
Psp124B I
PspE I
PspL I
PspN4 I
PspOM I
PspPP I
Pst I
Pvu II
Rsa I
Sal I
Sbf I
SfaN I
Sfi I
Sfr274 I
Sfr303 I
Sma I
Smi I
Sph I
Sse9 I
Ssp I
Tru9 I
Tth111 I
Vha464 I
Vne I
Vsp I
Xba I
Xma I
Zsp2 I
Not I
Aha III
Drd I
Hal I, Kpn49k I, Rsr I, Sso I
Ceq I, Eco32 I
Ehe I, Eco78 I
BssT1 I, EcoT14 I, Eco130 I, ErhB9 II, Sty I
None
Nde I
None
Ban II, Bsu1854 I, Bsp519 I, Bvu I, Eco24 I, Eco215 I, HgiJ II, SacN I
BsoF I, Bsp6 I, Fbr I, Fnu4H I, Ita I, Uur960 I
Bim19 II, Bsh I, BspK I, Bsp211 I, BsuR I, Dsa II, FnuD I, Pla I, Sbv I, Sfa I
BstF I, EcoV III, Hsu I, Ssb I
CviB I, FnuA I, Hha II
Bco27 I, BsiS I, Bst40 I, Hap II, Msp I, Sth134 I
HinP1 I, Hin6 I, SciN I
None
AtuC I, Bco102 I, BspX II, Fba I, Pov I
AspMD I, BspA I, Bsp105 I, Bsp143 I, BtK II, Dpn II, Mbo I, Nde II, Nla II, Sau3A I
None
Mch I, Nar I, Nda I, Nun II, SseA I
Eco56 I, NgoA IV, NgoM I
Asp700 I, BbuA I, Xmn I
Hpa II
Bme1390 I, Msp67 I, ScrF I
Bsp68 I, MluB2 I, Sbo13 I, Spo I
Ahd I, AspE I, Eam1105 I, EclHK I, Uba1190 I, Uba1191 I
BspC I, ErhB9 I, Nb1 I, Ple19 I, Pvu I, Rsh I, Xor II
Aat I, Eco147 I, SseB I, Stu I
Sac I, Sst I
Acr II, AspA I, BstE II, BstP I, Eca I, EcoO65 I, Eco91 I, NspSA II
BpuB5 I, BsiW I, Pfl23 II, PpuA I, Sp1 I,Sun I
AspN I, BscB I, Nla IV
Bsp120 I
Pfl27 I, PpuM I, Psp5 II
Api I, Asp713 I, BspB I, Bsp63 I, Hal II, Sfl I
Bav I, Dma I, Pvu84 II
Afa I
HgiC III, HgiD II, Nop I, Xci I
Sse8387 I
None
Sdi I
Abr I, Blu I, Ccr I, Mav I, PaeR7 I, Pan I, Sla I, Xho I, Xpa I
Cfr42 I, Csc I, Gal I, Kpn19 I, Ksp I, Sac II, Sst II
CfrJ4 I, PaeB I, PspAL I
Swa I
Bbu I, Pae I
TspE I, Tsp509 I
unfound
Mse I, Tru1 I
Asp I, Ats I
Afl II, Bfr I, BspT I, Bst98 I, Esp4 I, MspC I
Aaq I, Alw44 I, ApaL I, Sno I
Ase I, Asn I, PshB I, Vsp I
None
Ahy I, Cfr9 I, EaeA I, PspA I, Xcy I, XmaC I
EcoT22 I, Mph1103 I, Nsi I, PinB I, Sep I
GC GGCCGC
TTT AAA
GACNNNN NNGTC
G AATTC
GAT ATC
GGC GCC
C C(A/T)(A/T)GG
CCCGCNNNN
CA TATG
GGATG(9)
G(G/A)GC(C/T) C
GC NGC
GG CC
A AGCTT
G ANTC
C CGG
G CGC
GGTAC C
T GATCA
GATC
A CGCGT
GG CGCC
G CCGGC
GAANN NNTTC
C CGG
CC NGG
TCG CGA
GACNNN NNGTC
CGAT CG
AGG CCT
GAGCT C
G GTNACC
C GTACG
GGN NCC
G GGCCC
(G /A) GG(A/T)CC(C/T)
CTGCA G
CAG CTG
GT AC
G TCGAC
CCTGCA GG
GCATCNNNNN
GGCCNNNN NGGCC
C TCGAG
CCGC GG
CCC GGG
ATTT AAAT
GCATG C
AATT
AAT ATT
T TAA
GACN NNGTC
C TTAAG
G TGCAC
AT TAAT
T CTAGA
C CCGGG
ATGCA T
Enzymes
En
zyme
s
115
AATT ACGT AGCT ATAT CATG CCGG CGCG CTAG GATC GCGC GGCC GTAC TATA TCGA TGCA TTAA
Enzymes
En
zym
es
116
unit Cat No.
200 E-1111
1,000 E-1112
Source: Alcaligenes faecalis T2774.
Reaction Condition: AccuCut buffer Violet: 33 mM Tris-acetate, 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9,Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA,1 mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *10,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Ait I, Aor51H I, Fun I, Eco 47 III.
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: Unidentified
5' AGCGCT 3'3' TCGCGA 5'
unit Cat No.
200 E-1121
1,000 E-1122
Source: Alcaligenes faecalis T2774.
Reaction Condition: AccuCut buffer Violet: 33 mM Tris-acetate, 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9,Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *10,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Ait I, Aor51H I, Fun I, Eco 47 III.
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: Unidentified
5' AGCT...3'3' TCGA...5'
unit Cat No.
2,500 E-1141
12,500 E-1142
Source: Acetobacter pasteurianus.
Reaction Condition: AccuCut buffer Violet : 33 mM Tris-acetate, 10mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9, Incubate at 37 .
Storage Buffer : 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *20,000-100,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Ppe I.
Neoschizomer: PspOM I (GGGCCC).
Reactivity on methylated substrate
DNA: Blocked by overlapping dcm methylation (Cm5CWGG),
GGGCCm5C.
5' GGGCCC 3'3' CCCGGG 5'
unit Cat No.
, 1,000 E-2141
5,000 E-2142
Source: Vibrio species 343.
Reaction Condition: AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 8.5, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *40000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Ase I, Asn I, PshB I, Vsp I.
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: Unidentified
5' ATTAAT 3'3' TAATTA 5'
Enzymes
En
zyme
s
117
unit Cat No.
5,000 E-1211
25,000 E-1212
Source: Bacillus amyloliquefaciens H.
Reaction Condition: AccuCut buffer Orange : 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM KH2PO4, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *50,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: AccEB I, Ali I, Bna I, Bst I, NspSA IV, Sur I
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Not blocked by overlapping dam methylation Gm5ATC,
GGATCm5C, GGm6ATCC, GG m6ATCm5, GGATCm4C :
Blocked by GGA Tm4CC#, GGA tm5CC, GGAThm5Cm5C,
GGAhm5UCC.
5' GGATCC 3'3' CCTAGG 5'
unit Cat No.
1,000 E-1241
5,000 E-1242
Source: Bacillus globigii.
Reaction Condition: AccuCut buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1
mM 2- mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *20,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Ncr I, NspMAC I, Pae2k I, Pae18k I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Not blocked by GCm5CN5GGCb:
Blocked by Gm5CCN5GGC, GCCN5GGm5C GC m4CN5GGC.
5' AGATCT...3'3' TCTAGA 5'
unit Cat No.
200 E-2071
1,000 E-2072
Source: Streptomyces phaeochromogenes.
Reaction Condition: AccuCut buffer Orange : 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *5,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Bbu I, Pae I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by GCm6ATGC, Ghm5CATGhm5C. Not blocked by
GCATGm5C.
5' GCATGC 3'3' CGTACG 5'
unit Cat No.
2,000 E-1431
10,000 E-1432
Source: Bacillus stearothermophilus 2U.
Reaction Condition: AccuCut buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 60 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *20,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Aor I, Apy I, BseB I, Bse16 I, Bse17 I, Bse24 I,BspN I, Bst2 I, BstN I, BstO I, EcoR II,Fsp1604 I,Mva I, Sth117 I, Zan I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Not blocked by Cm5CWGG.
5' CC(A/T)GG 3'3' GG(T/A)CC 5'
Enzymes
En
zym
es
118
unit Cat No.
600 E-1271
3,000 E-1272
Source: Bacillus stearothermophilus 29.
Reaction Condition: AccuCut buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *15,000 units/m .
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Aag I, Ban III, Bci 29 I, Bsc I,Bsp106 I, Bsu15 I, Cla I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by Gm5ATC.
5' ATCGAT 3'3' TAGCTA 5'
unit Cat No.
400 E-1471
2,000 E-1472
Source: Bacillus stearothermophilus DE.
Reaction Condition: AccuCut buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 60 .
Storage Buffer: 10mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *25,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Dde I.
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: Unidentified
5' CTNAG 3'3' GANTC 5'
unit Cat No.
2,000 E-1601
10,000 E-1602
Source: Deinococcus radiophilus.
Reaction Condition: AccuCut buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1mM EDTA, 50%glycerol, pH 7.5, 1 mM 2-mercaptoethanol, Store at -20
Concentration: *10,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Aha III.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by TTTAm6AA
5' TTTAAA 3'3' AAATTT 5'
unit Cat No.
200 E-1781
1,000 E-1782
Source: Kurthia zopfil 9.
Reaction Condition: AccuCut buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *5,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: AspMD I, BspA I, Bsp105 I, Bsp143 I, BtK II,Dpn II, Mbo I, Nde II, Nla II, Sau 3A I
Neoschizomer: Dpn I (GATC).
Reactivity on methylated substrate
DNA: Not blocked by Gm5ATC.
5' GATC 3'3' CTAG 5'
Enzymes
En
zyme
s
119
unit Cat No.
5,000 E-1621
25,000 E-1622
Source: Escherichia coli.
Reaction Condition: AccuCut buffer EcoR1: 100 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 20 mM Tris-HCl, 300 mM KCl, 1 mM EDTA, 10mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *20,000-100,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Hal I, Kpn49k I, Rsr I, Sso I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by Gm6ATTC, GA m6ATTC, GAAT Tm5C. Not
blocked by GAATThm5C, GAAhm5Uhm5UC.
unit Cat No.
2,000 E-1631
10,000 E-1632
Source : Escherichia coli.
Reaction Condition : AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 8.5, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 300 mM KCl, 0.1 mM EDTA, 10mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *20,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Ceq I, Eco 32 I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by Gm6ATATC, GA Tm6ATC. Not blocked by GATA
Tm5C, GATAThm5C.
5' GAATTC 3'3' CTTAAG 5'
5' GATATC 3'3' CTATAG 5'
unit Cat No.
50 E-1661
250 E-1662
Source: Flavobacterium aquatili.
Reaction Condition: AccuCut buffer Green: 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 55 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *500 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: None .
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: Unidentified
5' CCCGCNNNN 3'3' GGGCGNNNNNN 5'a
unit Cat No.
3,000 E-1711
15,000 E-1712
Source: Haemophilus aegyptius.
Reaction Condition: AccuCut buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *20,000 units/ml.
Heat Inactivation: No.(65 for 20 minutes)
Isoschizomer: Bim19 II, Bsh I, BspK I, Bsp211 I,BsuR I, Dsa II, FnuD I, Pla I, Sbv I,Sfa I
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by GGm5CC. Not blocked by GGCm5C.
5' GGCC 3'3' CCGG 5'
Enzymes
En
zym
es
120
unit Cat No.
2,000 E-1731
10,000 E-1732
Source: Haemophilus influenzae.
Reaction Condition: AccuCut buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *20,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: CviB I, FnuA I, Hha II.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by Gm6ANTC, GANThm5C. Not blocked by GANT m5C.
5' GANTC 3'3' CTNAG 5'
unit Cat No.
1,000 E-1161
5,000 E-1162
Source: Arthrobacter species LE3860
Reaction Condition: AccuCut buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercapto ethanol, 50% glycerol pH 7.5, Store at -20 .
Concentration: *20,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Cfo I, Fnu D III, Hha I.
Neoschizomer: HinP I (GCGC), HspA I (GCGC).
Reactivity on methylated substrate DNA: unidentified.
5' GCGC...3'3' CGCG...5'
unit Cat No.
10,000 E-1721
50,000 E-1722
Source: Haemophilus influenzae Rd.
Reaction Condition : AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 8.5, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 300 mM Nacl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% 50% glycerol, pH 7.5,Store at -20 .
Concentration: *20,000-100,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: BstF I, EcoV III, Hsu I, Ssb I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by AAGhm5CTT, AAGm5CTT, m6AAGCTT. Not
blocked by AAGChm5Uhm5U, A m6AGCTT.
5' AAGCTT...3'3' TTCGAA...5'
unit Cat No.
200 E-1831
1,000 E-1832
Source: Moraxella species.
Reaction Condition: AccuCut buffer Green : 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *15,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Hpa II.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by m5CCGG, m5CCGG, m5Cm5CGG. Not blocked bym4CCGG, Cm4CGG, Cm5CGG.
5' CCGG 3'3' GGCC 5'
Enzymes
En
zyme
s
121
unit Cat No.
1,000 E-1741
5,000 E-1742
Source: Haemophilus parainfluenzae.
Reaction Condition: AccuCut buffer Green: 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *5,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Bco27 I, Bsi S I, Bst40 I, Hap II, Msp I, Sth134 I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by m4CCGG, m5CCGG, Cm4CGG, Cm5CGG,hm5Chm5CGG.
5' CCGG 3'
3' GGCC 5'
unit Cat No.
3,000 E-1761
15,000 E-1762
Source: Klebsiella pneumonia.
Reaction Condition: AccuCut buffer Green: 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *15000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: None.
Neoschizomer: Acc65 I, Asp718, Asp718 I (GGTACC)
Reactivity on methylated substrate
DNA: Blocked by GG Tm6ACC, GGTAm4CC, GGTAm5Cm5C,
GGTACm4C. Not blocked by GGTAm5CC, GGTACm5C
5' GGTACC 3'3' CCATGG 5'
unit Cat No.
1000 E-1791
5,000 E-1792
Source: Micrococcus luteus.
Reaction Condition: AccuCut buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *20,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: None.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by Am5CGCGT. Not blocked by m6ACGCGT.
5' ACGCGT 3'3' TGCGCA 5'
unit Cat No.
200 E-2101
1,000 E-2102
Source: Thermus ruber 9.
Reaction Condition: AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM KCl, 1 mM DTT, pH 8.5, Incubate at 65 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *20,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Mse I, Tru1 I.
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: Unidentified
5' TTAA...3'3' AATT...5'
Enzymes
En
zym
es
122
unit Cat No.
200 E-1801
1,000 E-1802
Source: Micrococcus lylae 113.
Reaction Condition: AccuCut buffer Green: 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *5,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Mch I, Nar I, Nda I, Nun II, SseA I.
Neoschizomer: Bbe I (GGCGCC), Ege I, Ehe I (GGCGCC), Kas I (GGCGCC)
Reactivity on methylated substrateDNA: Unidentified
5' GGCGCC 3'3' CCGCGG 5'
unit Cat No.
500 E-1671
2,500 E-1672
Source: Flavobacterium aquatili ND.
Reaction Condition: AccuCut buffer Violet : 33 mM Tris-acetate, 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9,Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *10,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Nde I.Neoschizomer: Unfound
Reactivity on methylated substrate DNA: Unidentified
5' CATATG 3'3' GTATAC 5'
unit Cat No.
500 E-1381
2,500 E-1382
Source: Bacillus species 19.
Reaction Condition: AccuCut buffer Blue: 10 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 8.5, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *5,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Nco I.
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: unidentified
5' CCATGG 3'3' GGTACC 5'
unit Cat No.
600 E-1201
3,000 E-1202
Source: Actinobacillus suis NH.
Reaction Condition: AccuCut buffer Green: 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *30,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Nhe I, Pst NH I.
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: unidentified
5' GCTAGC 3'3' CGATCG 5'
Enzymes
En
zyme
s
123
unit Cat No.
10,000 E-1951
50,000 E-1952
Source: Providencia stuartii.
Reaction Condition: AccuCut TM buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *50,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Api I, Asp713 I, BspB I, Bsp63 I, Hal II, Sfl I.
Neoschizomer: Unfound
Reactivity on methylated substrateDNA: Blocked bym5CTGCAG/m5CTGm5CAG/CTGm5CAC/CTGCm6AG
unit Cat No.
1,500 E-1961
7,500 E-1962
Source: Proteus vulgaris 84.
Reaction Condition: AccuCut TM buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *30,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Bav I, Dma I, Pvu84 II.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by m5CAGCTG/CAGm5CTG/CAGm4C T G
5' CTGCAG 3'3' GACGTC 5'
5' CAGCTG 3'
3' GTCGAC 5'
unit Cat No.
200 E-1591
1,000 E-1592
Source: Curtobacterium citreus N.
Reaction Condition: AccuCut buffer Violet: 33 mM Tris-acetate,10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9, Incubateat 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *5,000 units/ml. (Assayed Adenovirus-2 DNA)
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Not I.
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: Unidentified
5' GCGGCCGC 3'3' CGCCGGCG 5'
unit Cat No.
3,000 E-2031
15,000 E-2032
Source: Streptomyces fradiae 274.
Reaction Condition: AccuCut buffer Green: 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 50 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *10,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Abr I, Blu I, Ccr I, Mav I, PaeR7 I, Pan I, Sla I, Xho I, Xpa I.
Neoschizomer: Unfound
Reactivity on methylated substrate DNA: Unidentified
5' CTCGAG 3'3' GAGCTC 5'
Enzymes
En
zym
es
124
unit Cat No. 5'. GCATCNNNNN 3'3'. CGTAGNNNNNNNNN 5'
Source: Streptococcus faecalis N.
Reaction Condition: AccuCut buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *1,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: None.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Not blocked by GCATm5C.
unit Cat No.
1,000 E-1971
5,000 E-1972
Source: Rhodopseudomonas sphaeroides.
Reaction Condition: AccuCut buffer Green: 10 mM Tris-HCl,10 mM MgCl2, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *10,000 units/ml.
Heat Inactivation: No. (65 for 20 minutes)
Isoschizomer: Afa I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by GTm6AC /GTA m4C. Not blocked by GTAm5C.
5' GTAC 3'3' CATG 5'
unit Cat No.
1,500 E-1891
7,500 E-1892
Source: Pseudomonas species 124B.
Reaction Condition: AccuCut buffer Orange: 10 mM Tris-HCl,10 mM MgCl2, 50 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *15,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Sac I, Sst I.
Neoschizomer: Ecl136 II, EcoICR I (GAGCTC).
Reactivity on methylated substrate DNA: Unidentified
5' GAGCTC 3'3' CTCGAG 5'
unit Cat No.
1,000 E-1981
5,000 E-1982
Source: Streptomyces albus.
Reaction Condition: AccuCut buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT pH 7.6, Incubate at 37 .
Storage Buffer: 10mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1 mM2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *20,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: HgiC III, HgiD II, Nop I, Xci I.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by G tm5CGAC, GTCGm6AC, Ghm5UCGAC. Not
blocked by GTCGAm5C.
5' GTCGAC 3'3' CAGCTG 5'
50 E-2001
250 E-2002
Enzymes
En
zyme
s
125
unit Cat No.
1,000 E-2051
5,000 E-2052
Source: Serratia marcescens.
Reaction Condition : AccuCut buffer Violet: 33 mM Tris-acetate, 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9,Incubate at 25 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration :*25,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: CfrJ4 I, PaeB I, PspAL I.
Neoschizomer: Cfr9 I, Xma I (CCCGGG)
Reactivity on methylated substrate
DNA: Bolcked by m4CCCGGG, m5CCCGGG, Cm4CCGGG,
CCm4CGGG, CCm5CGGG. Not blocked by Cm5CCGGG.
5' CCCGGG 3'3' GGGCCC 5'
unit Cat No.
2,000 E-2151
10,000 E-2152
Source: Xanthomonas badrii.
Reaction Condition: AccuCut buffer Red: 50 mM Tris-HCl, 10mM MgCl2, 100 mM NaCl, 1 mM DTT, pH 7.6, Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20 .
Concentration: *25000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: None.
Neoschizomer: Unfound
Reactivity on methylated substrate
DNA: Blocked by TCTAGm6A, Tm5CTAGA, Thm5CTAGA.
5' TCTAGA 3'3' AGATCT 5'
unit Cat No.
50 E-2161
250 E-2162
Source: Xanthomonas malvacearum
Reaction Condition: AccuCut buffer Violet: 33 mM Tris-acetate, 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT, pH 7.9,Incubate at 37 .
Storage Buffer: 10 mM Tris-HCl, 50 mM KCl, 0.1 mM EDTA, 1mM 2-mercaptoethanol, 50% glycerol, pH 7.5, Store at -20
Concentration: *2,000 units/ml.
Heat Inactivation: Yes. (65 for 20 minutes)
Isoschizomer: Ahy I, Cfr9 I, EaeA I, PspA I, Xcy I, XmaC I.
Neoschizomer: Sma I
Reactivity on methylated substrate DNA: Unidentified
5 CCCGGG 33 GGGCCC 5