L22 Flashcards
(17 cards)
how long is a primer in a PCR
~20 nt
what enzyme is needed for PCR
heat stable Taq polymerase
PCR stands for
polymerase chain reaction
how do primers bind target
cool down tube
how many cycles does PCR need to make the right size of DNA
three cycles
what is added last in a PCR test
taq DNA polymerase
what are the 4 things in PCR
primers
heat stable polymerase
dNTPs
template DNA
where does specificity of copying arise from in PCR
primers that bind unique sequences
what becomes limiting in PCRs
enzyme
Tm
temperature that a double stranded sequence of the primer would fully melt into single stranded molecules
what is annealing temp
when the primers bind to the template DNA
how much lower is annealing temperature than Tm of primer
~5 degrees
positive control
having a working template from another sample to compare
whats a negative control in PCR
entire reaction but with no template DNA to see if any contamination occurred
basic PCR
every tube cycles together
gradient PCR
allows optimization of annealing temps
real time PCR
allows quantification in each cycle
