Basics of tics
Transcript of Basics of tics
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Dr: A. Ellethy
College of Dentistry
Lec. 2
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What, why, when, where study gene?
How to design a primer?
Types of practical used primers
How to read DNA sequences (the most recently
developed program in USA).
Data analysis for sequenced DNA
Top secretes to study DNA sequences
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What,
why,
when,
where
study
gene?
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WHERE IS DNA? WHY?
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Under sterile conditions
Automated DNA sequencer
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PCR MIX Volume
H2O 12 ul
5X Q soln 5
10X buffer 2.5
50X dNTPs 0.5
primer A 1
primer B 1
Taqpolymerase 0.5
To amplify DNA sequences?
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Labeled ddNTPs with
Radioisotopes S35
Fluorescentlylabeled
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1 cycle: 97C 2 min
30 cycles:
Denaturation 97C 1 min
Annealing XC 1 minExtension 72C 1min
1 cycle:
72
C 6min
store:
4C store
X based melting temperature: 55-60 C
PCR PROGRAME
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A G C T
Start to read DNA sequences
HOW TO READ RADIOGRAPHIC SHEETS OF DNASEQUENCES?
2 hours run 1
4 hours run 2
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TGCTTATTACTGCA
GTCACCATC
-------------------?
A G C T
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DNACHROMAS
Open and save sequences in Applied Biosystems, Staden Chromatogram (SCF), FASTA,EMBL, GenBank, SwissProt, GenPept, GCG RSF and plain text formats.
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HOW TO USE DNA CHROMAS?
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Cell Chromosome DNACell Chromosome DNA
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DNA SEQUENCE
ANALYSIS
IDENTIFICATION
CONFIRMATION
APPLICATION
DIAGNOSIS
VACCINATION
REPAIRING
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ATGGTCTTATTAAGAAAAAACTTCTGTTGTACCATAGGTTATGCCCATATTTCCTCCACCACAACTTCAA
ATCCTTCATCAGAAACGAAGAGCATAGCCACAGACTATGAAACCACCCCGACCACCACTATATCTAGCAC
TGCCTCAGGATCTACGCCCACCCTGACCACCACTGCATCCAGCATTGCCTCAGACTCTAAGCCCACCCTG
ACCACCACTGCATCCAGCACTGCCTCAGGATCTATGTCCACCCCGACCACCCCTGCATCCAGCACTGCCT
CAGGCTCTACACCCACAACGACCACCACTGCATCCAGCACTGCCTCAGGATCTACACCTACCCCGACCAC
TACTGCATCTAGCAAAGCCTCAAGATCTGTGCCCACCACTGTATCCAGCACTGGCTCAGGCTCTACACCC
ACTCCGACCACCACTGCATCCAGCACTGCCTCAGGATCTACGCCCACCCTGACCACCACTCCATCCAGCA
CTGCCTCAGGATCTACACCTACCCCGACCACCCCTGCATCCAGTACTGCCTCAAGCTCTTCACCCACCCC
AACCACCCCTGTATCCAGCACTGCCTCAGGATCTACACCCACCCCGACCACCACTGCATCTAGCAAGGCC
TCAGGATCTGTGCCCACCACTGTATCCAGCACTGGCTCAGGCTCTACACCCACTCTGACCACCACTGTAT
CCAGCACTGTCTCAGACTCTACACCCACCCCGACCACCACTGCATCCAGCACTGCCTCAGGATCAGCACC
CACCCCGACCACCACTGCATCTAGCACTGCCTCAGGATCTATGCCCACCCTGACCACCAATGCATCCAGC
Dentin Sialophosphoprotein EST
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Put original DNA sequences
Put original test sequences
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Blastn dentin sequences
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)1430-1431(
150 50
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100 .