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In using the polymerase chain reaction (PCR), briefly explain?
a. Why two primers and not one or more than two are used
b. why the reaction must be heated to more than about 80 degrees at regular intervals
c. why the enzyme that copies the DNA sequence is not inactivated by this high temperature
d. why PCR primers used to score mini or micro-satellite polymorphism ( simple sequence repeats (SSR) do not include the repeat sequence itself
A. DNa is synthesized 5'-3'. There are two strands, and since each goes 5'-3', two primers are used, one for each strand, that gets places on the 5' side.
B. The reaction is heated because this denatures the DNA, or breaks the strands apart, so that the primers and bases can be added to the template to create a new DNA strand.
C. The enzyme typically used for this is Taq polymerase, which has an optimal temperature that is very high (I can't think of the temperature off the top of my head right now) But this enzyme has been found in the hot springs, like Old Faithful and stuff, it like hot environments
D. If you have a primer that has in it something that repeats, the chance of the primer settling somewhere that we don't want it to be is much higher. Primers need to be specific so that the person analyzing the DNA can be sure of exactly what they are looking at.
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