Lecture 3 Flashcards
1
Q
What used to be the method of quant after amplification and how would it work?
A
SLOT BLOT:
primate specific biotinylated oligo probe that binds 2 specific region (D17Z1 satellite locus)
2 nylon sheets. cut slots into one . put on top. she falls and cant move. probe flood.
2
Q
What is often used to link DNA and other molecules?
A
Biotin!
3
Q
What was the issue w slot blot
A
- low sensitivity
aka false negative
4
Q
DNA replication in vivo
A
- synthesized in the 5’ to 3’
- add nucleotides to the 3’ end
5
Q
What r the components of PCR
A
- DNA pol (Taq)
- dNTPs
- oligo primers
- MgCl2 (counters neg charge of DNA phosphate backbone)
- (Bovine serum albumin)
6
Q
What technical innovation made pcr easy?
A
- TAQ
- Thermocycler
7
Q
WHat did the primitive PCR process include?
A
E. coli Pol
water baths
8
Q
Who invented PCR
A
Kary Mullis
9
Q
What are the typical PCR temps
A
denaturation :94
annealing 35-60
ext: 72
10
Q
What is the most efficient distance betwn the primers
A
<3 kb
