Question

1. a. If you are asked to amplify exon 3 of beta hemoglobin gene how you would do this? (points: 6)

b. PCR revolutionized molecular biology-explain. (points: 4)

Answer 1

A ) Exon 3 of beta hemoglobin gene can be amplified via PCR

1 The genomic DNA can be amplified in two different PCR master mixes

2 PCR I with ddH2O, 5 U of TaqGold ,PCR amplification buffer, magnesium chloride ,4dNTP mix, primer A, primer B, primer D, DNA.

3 PCR II with the same reagents except for the primers, primer C and primer D.

4 35 cycles of denaturing at 95°C can be done using thermocycling process , annealing at 58.5°C for 1 minute, and extension at 72°C for 2 minutes can be done . Final extension at 72°C for 10 minutes can take place .

5 The amplification products were then can be subjected to electrophoresis in agarose gel.

6 At the end the amplicons can be purified and sequenced .

b) The polymerase chain reaction (PCR) revolutionized the field of molecular biology

The small size dna can be amplified million times using PCR , which can be used for further analysis .

Steps includes :

1 Denaturation of the template DNA via heating

2    Annealing

3   Extension

This process is done in thermocycler using DNA Taq polymerase

Using PCR a specific sequence can be amplified fastly .

PCR can be used to detect pathogens that are difficult to culture

PCR can be used for cancer diagnosis.

It plays vital role in Forensic studies

It has more applications in the field of molecular biology and medicine .

I love’d to hear your feedback and Rating. If you like the answer please give me a positive rating, which will boost me more. Thank you.