(2pts) Basic steps to clone a gene into a vector for expression: (1)PC Ramplification of the gene, (2) digest (cut) both the double-stranded PCR product (insert) and plasmid vector with the same restriction enzyme(s) to create complementary “sticky ends”, (3) allow the digested insert and vector to anneal, and (4) ligate the annealed fragments together with DNA Ligase. A. What additional step is required to generate a DNA template for PCR from a eukaryotic organism rather than a prokaryotic organism, and why? B. Why is it necessary to re-circularize (ligate) a plasmid prior to transformation into cells?
Basic steps to clone a gene into a vector for expression
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