1
Q
Library Cloning
A
- cDNA is cleaved and put in vectors
- each vector has a different piece of DNA and these are put in bacteria
- plated on petri plates
2
Q
Why convert mRNA to cDNA
A
more stable
3
Q
why use cDNA lib instead of mRNA
A
when you use cDNA you only look at expressed genes
4
Q
how to convert RNA to cDNA
A
- isolate the mRNA via poly-A-tail
- reverse transcriptase uses mRNA as a primer to make DNA strand
- Have an RNA-DNA hybrid
- RNaseH cuts the RNA
- DNA pol1 and DNA ligase synthesizes the second DNA strand
- now there is DS DNA
5
Q
Sanger Sequencing
A
-DS DNA molecule
Bio 230
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