6. In PCR, the purpose of dNTPs is to a. Cleave the DNA into small fragments b.

Question

6. In PCR, the purpose of dNTPs is to a. Cleave the DNA into small fragments b. Increase the temperature of the reaction c. Serve as DNA primers providing a free 3'-OH end for DNA synthesis d. Add nucleotides to the growing DNA strand Taq polymerase is a DNA polymerase enzyme that was originally isolated from Thermus aquaticus bacterium. It is used in PCR because 7. a. It can withstand high temperatures b. It can withstand high pressure c. It can polymerize DNA a lot faster than any other type of enzymes 8. In gel electrophoresis, smaller fragments move through the gel a. b. c. Slower Faster At the same rate as the large fragments 9. Which of the following techniques is used for visualizing 300 bp fragment PCR product? a. Southern Blotting b. Fluorescent labeling with green fluorescent protein (GFP) c. Polyhistidine tagging d. Polyacrylamide gel electrophoresis e. Agarose gel electrophoresis

Explanation / Answer

PCR is used to amplify target gene sequences. It is an invaluable technique used in molecular biology and genetics.

PCR reaction components:

Taq polymerase

Buffer

iii. dNTPs

iv. Primers

v. Template

PCR steps:

i. Denaturation (92'C to 94'C): Unwinding of the DNA duplex

ii. Annealing (50'C to 65'C): Binding of the primer to the template DNA

iii. Extension (70'C to 72'C): Extension of the primer by Taq polymerase

iv. Repeat

6. Option D is correct.

dNTPs serve as substrates during PCR reaction.

7. Option A is correct.

Taq polymerase can withstand high temperature.

8. Option B is correct.

Smaller fragments run faster during gel electrophoresis.

9. Option E is correct.

We can use agarose gel electrophoresis to visualize a 300 bp DNA fragment.

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