Question

1. Using your understanding of PCR amplification answer the following questions:

a) How do you know if a primer dimer is created during PCR?

b) What is nonspecific PCR amplification and how do you know if it exists during PCR?

c) What are the possible results you could expect to observe on the agarose gel from your PV92C results? Use a schematic diagram and draw a gel to illustrate where the PV92 primers bind. You should also consider where the primers bind on the paternal and maternal chromosomes as well.

Answer 1

Answer1)- A):-

:- Primer Dimer is a product of PCR that is formed when the complementary strands of primer DNA binds with each other.

It can be recognised by the following methods:-

• In results of gel electrophoresis :- When we stain the PCR product using Ethidium Bromide then if Primer Dimer would have been formed then it is seen like a band at around 30-60 base pairs with good intensity while the actual target sequence is located at a much higher base pair location.
• Using Melting Curve Analysis :- The denaturing Temperature of a Primer Dimer is very low owing to its very short sequences, while it is opposite for the target sequence. Hence we can easily identify either of them.

(According to the Chegg guidelines I am unable to answer all the questions, hope you understood this one very well !)