2. Subcloning Primer design is an essential skill for successful PCR amplificati

Question

2. Subcloning Primer design is an essential skill for successful PCR amplification. Let say that you want to repeat this semester's subcloning. Using the given coding sequence, choice of enzymes and vector, (a) design a forward and reverse primers to amplify the given coding sequence keeping in mind that you are planning to subclone it into the given vector (/2 Marks). (b) Explain how the forward primer was (or wasn't) modified to keep the reading frame of the amplicon in frame with the reading frame of the vector. ( /2 Marks)

Explanation / Answer

a) PMID- 22426482

b) Sequencing of T3 RNA polymerase- 2989096

1985;33(3),351-5

Cloning of T3 RNA polymerase- 3011596

1986;41(2-3),193-200

c) PMID- PMC46256

N748 at -8

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