I Ý asnc4u.co.kr/upload/editordata/PCRGUIDE_ALL.pdf · 2018-06-01 · 1$3 i Æ î i ä s À ² @...
Transcript of I Ý asnc4u.co.kr/upload/editordata/PCRGUIDE_ALL.pdf · 2018-06-01 · 1$3 i Æ î i ä s À ² @...
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TaKaRa Gradient ThermalCycler Dice
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The Best PCR EnzymesThe Best PCR Enzymes
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HOOH
HO
HO
OH
OH
HO OH
OH
HO
PP
HOOHP
P
HO
HO
OH
OH
3’
5’3’
5’
A3’
5’
5’
3’A
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CGC
GGATCC
5’
3’
3’
3’
5’
5’
3’
PCR5’
CTTAAGGC
GAATTCCGCTTAAGGC
CGCGGATCCGCGCCTAGG
3’
5’
5’
3’
GCTTAAp
pAATTCCGGGC
pGATCCG
GAATTC
CCTAGG
CGCGGCGCCTAGp
Vector
5’ CGCGGATCC 3’ 5’ CGGGATCC 3’
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PPP
OH
C
G
P
A
P
T
P
C
P
A
P
T
P
C
P
A
P
T
P
C
P
A T
OH
PPP
C
G
P
A
P
T
P
C
P
A
P
T
P
C
P
A
P
T
P
C
P
A T
P
HO
C
P
G
P
A
P
T
P
P
A
P
C
P
G
P
A
P
T
P
P
A
P
C
P
G
P
A
P
T
P
P
A
P
C
P
G
P
A
P
T
P
P
A
P
HO
P
HO
C
P
G
P
A
P
T
P
P
A
P
C
P
G
P
A
P
T
P
P
A
P
C
P
G
P
A
P
T
P
P
A
P
C
P
G
P
A
P
T
P
P
A
P
HO
P
G
P
T
P
A
P
G
P
T
P
A
P
G
P
T
P
A
P
G
P
T
P
A
P
P
HO
C
P
G
P
A
P
T
P
U
P
A
P
C
P
G
P
A
P
T
P
U
P
A
P
C
P
G
P
A
P
T
P
U
P
A
P
C
P
G
P
A
P
T
P
U
P
A
P
HO
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P
T
P
T
HO
PP
P
P
P P
OHPP
P
P
P P
OH
HO
PP
P
P
P P
P
A
HO
P
P
P
P P
HO
P
A
P
P
P
P P
OHPP
P
P
P P
P
T
PP
P
P
P P
PPP
T
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5 l
10 l
20~50 l
μμ
μ
μ
μ
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GGATCC
CCTAGG
5’
3’
5’
3’
GATATC
CTATAG
5’
3’
5’
3’
GGATCC
CCTAGG
5’
3’
5’
3’
GATATC
CTATAG
5’
3’
5’
3’
GGATCA
CCTAGG
5’
3’
5’
3’
GGATCAATC
CCTAGTTAG
5’
3’
5’
3’
GATATC
CTTTAG
5’
3’
5’
3’
GCGGCCGC
CGCCGGCC5’
3’
5’
3’
CAGCTG
GTCGAC5’
3’
5’
3’
GCGGCCGC
CGCCGGCC5’
3’
5’
3’
GATCTG
CTAGAC
5’
3’
5’
3’
GCGGCCCC
CGCCGGCC
5’
3’
5’
3’
GCGGCCCCTG
CGCCGGGGAC
5’
3’
5’
3’
GATCTG
CTGGAC
5’
3’
5’
3’
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Real Time Thermal Cycler
ö
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AB1B0B
2 k
(bp)
710
489
404
325
GH
CDEF
IJKL
MN
OP
Q
R
S
N N T T
Case 1
N N T T
Case 2
N N T T
Case 3
N N T T
Case 4
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“ ”
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’
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m/m
m/+ m/+
m/m +/+
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è
ü
è
ü
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TaKaRaPCR Thermal Cycler
MP TP3000
Sample Block forPCR Tube Plate
Sample Block for SlideGlass(in situ PCR)
Sample Block for0.5 ml 54-well
Sample Block for1.5 ml 30-well
TP3000Sample block
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x
y
x
y
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TTTTTTAAAAAA
TTTTTTAAAAAA
TTTTTTAAAAAA
AAAAAA
TTTTTT
TTTTTT
AAAAAATTTTTT
AAAAAATTTTTT
5’
5’3’
5’3’
5’3’
5’3’
3’
3’
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AAAAAAAA
AAAAAAAA
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·
··· · ···· ·· ····· ·
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PCRWITH-TAQANDPRIMERSAMPLIFIEDASPECIFICDNAFRAGMENT
PCRWITHREVERSETRANSCRIPTASEDETECTEDANR--NAFRAGMENT
PCRWITHDEGE-NE-RATEPRI-MERSGIVESMEANEWCDNAFRAGMENT
******* * * **********
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A A'B C C'
A A’
B
C C’
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AAAAAA
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’ ’
’ ’
’ ’
5'
5'
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AAAAAAmRNA
GSP2 GSP1
GSP2 GSP1
GSP2
Adaptor
Adaptor
Adaptor
GSP1
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5’ - CUA CUA CUA CUA GGC CAC GCG TCG ACT AGT ACG GGI IGG GII GGG IIG – 3’
’ Mlu I Spe I
Sal IUDG cloning site
5’ - CUA CUA CUA CUA GGC CAC GCG TCG ACT AGT AC – 3’
Mlu I Spe I
Sal IUDG cloning site
GGC CAC GCG TCG ACT AGT AC – 3’
Mlu I Spe I
Sal I
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N T1 T2 N T
10 11
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N T1 T2
2
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300-
250-
200-
150-
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