a) You have to run an important pcr reaction. Unfortunately you can\'t find any

Question

a) You have to run an important pcr reaction. Unfortunately you can't find any TAQ in the whole department. The only DNA poly DNA polymerase that is available is the Klenow fragment (E. coli DNA polymerase without exonuclease activity). Can you still run your pcr reaction using this DNA polymerase? Give a detailed explanation!

b) You made a 1% agarose gel for the DNA fingerprinting experiment whereas a 2% agarose gel for this experiment. What could be the reason for it? Give a detailed explanation.

Explanation / Answer

(a) you cannot run your pcr reaction using this DNA polymerase since this is not heat-stable. The Taq DNA Polymerase is a thermostable enzyme that synthesizes DNA from single-stranded templates in the presence of dNTPs and a primer. Since there is thermocycling step in the PCR where the temperature is raised to 90C, the EColi DNA polymerase could denature and could be rendered useless.

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