LECTURE: Methods and Membrane Structure Flashcards
What is PCR useful for?
Getting a bunch of copies of a DNA segment
Why would you use PCR to make cDNA?
cDNA doesn’t have introns
How could PCR diagnose diseases?
You can get blood and stick primers for e.g. virus DNA in it, then amplify. See if you have a result
PCR: Why do you heat first?
Separate the two DNA strands
PCR: Why do you cool after heating?
Attaches the primers on to DNA segment
What do you need to add to PCR first to make cDNA? What enzyme is used?
mRNA sequence. You use reverse transcriptase to copy it
How do you terminate your sequencing in DNA sequencing? Why?
Add ddNTPP’s. They don’t have 3’ OH so can’t polymerize
How do you terminate your sequencing in DNA sequencing? Why?
Add ddNTPP’s. They don’t have 3’ OH so can’t polymerize
How does manual sequencing work?
Get 4 tubes and PCR.
Add chain-terminating ddNTPs for specific nucleotides
(e.g. A > ATGTCA, etc)
What’s the difference between automated and manual sequencing?
Automated: send your sample to a company, they do it for you.
-Fluorescently labeled nucleotides
What direction is 5’ in sequencing gels?
Down (negatively charged side)
Why is an expression vector useful?
It’s a box to put the gene you want to express into
What do you use to cut DNA after you put it in the promoter?
Restriction nuclease
