QUESTION 6 3 marks) You are interested in a particular segment of DNA and would

Question

QUESTION 6 3 marks) You are interested in a particular segment of DNA and would like to insert it into a cloning vector. You have isolated your "DNA of interest" from a DNA library, and have identified all the restriction enzyme recognition sites in the region of your "DNA of interest" EcoRI Hindll Xbal Sph coRI Hind Sph Hindl Not DNA of Inter You plan to clone your "DNA of interest" into the following cloning vector. EcORI Not Ampicillin resistance gene Xbal Multiple Cloning Site Hindlll Promoter Origin of Replication

Explanation / Answer

1. DNA of interest should be treated with XbaI and NotI because using another combination may causevfragmentation of DNA of interest.

2. Primer used in PCR are of two type forward and reverse primer which acts on complimentary strands. Primer 5'GTTC3' is used as forward primer and 5'GCCC3' is used as reverse primer to produce amplified PCR product.

3. Complimentry strand sequence should be exactly similar to ddNTPs obtained. Hence-

5'TGACACGT3'

And templete strand should be-

5'ACGTGTCA3'.

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