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Oct 14, 2023

Question: Cloning Primer design is a skill essential to the success of a PCR reaction. Let`s say you want to repeat the subcloning we did this semester. Using the sequence from your section, choice of restriction enzymes and vector,

(a) please design a forward and reverse primer to amplify your sequence keeping in mind that you must use the given vector.

(b) Explain how the sense primer was (or was not) modified to fit the reading frame of the amplicon into the reading frame of the

The gene coding sequence for your section can be downloaded fromthe NCBI website. A photo of the multiple cloning site for the differentvectors is available on the course`s Brightspace site (Hypo Vectors inthe Lab 2 section). The first enzyme must be used for the sense primer whilethe second will be used to design the antisense primer. Use 20 bp of codingsequence for your two primers.

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