DNA Sequencing Flashcards
1
Q
What are the steps of automated Sanger DNA sequencing?
A
- Reaction mixture prep
- In vitro DNA synthesis
- Capillary electrophoresis
- Data interpretation
2
Q
What is in the reaction mixture for automated Sanger DNA sequencing?
A
Template DNA One oligo primer Taq DNA polymerase Lots of normal dNTPs Some labelled ddNTPs
3
Q
How do we do in vitro DNA sequencing for automated Sanger DNA sequencing?
A
Use a standard PCR machine
25 cycles of denature/anneal/extend
4
Q
What is the difference in shape between PCR and automated Sanger DNA sequencing?
A
PCR = dsDNAs of equal length
Automated Sanger DNA sequencing = ssDNAs of unequal length
5
Q
what is the difference in number between PCR and Automated Sanger DNA sequencing?
A
PCR = 1 billion products per template
Automated Sanger DNA sequencing = 25 products per template
6
Q
What is the max size difference between PCR and Automated Sanger DNA sequencing?
A
PCR = 5 kb
Automated Sanger DNA sequencing = 700 nt
