Question

What is it called when a PCR technician determines the best conditions for running a PCR...

What is it called when a PCR technician determines the best conditions for running a PCR protocol?

Describe the function of each of the following PCR components: dNTPs, reverse primers, forward primers and reaction buffer.

Give 3 examples where PCR is used.

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Answer #1

1. OPTIMIZATION refers to the process of finding out best conditions for a PCR reaction to occur.

It includes changing one or all of the following factors.

i. Primer concentration

ii. Annealing temperature

iii. Template concentration

iv. Mg+2 concentration

v. DMSO concentration

2.

dNTPs = Building blocks of new strands

Reverse and forward primer: They bind to teh template and get extended by the Taq polymerase

Reaction buffer provides optimal salt concentration and required pH for the reaction to occur.

3.

Uses of PCR

1. Quantification of a transcript (RT PCR)

2. Genotyping

3. Cloning a gene

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