You are given 3 tubes of RNA and asked by your supervisor to clone the gene encoding the proinsulin C-peptide that will be used later to produce insulin. The first tube contains purified mRNA isolated from pancreatic β-cells from a patient in a postabsorptive state. The second tube contains purified mRNA isolated from pancreatic α-cells from a patient during absorptive state. The third tube contains purified mRNA isolated from pancreatic β-cells from a patient during absorptive state. In not more than 2 pages (using 1.5 line space of Arial or Times New Roman fonts) provide answers for the following questions? 1) Which tube from the three is the most appropriate to use and why? 2) Describe the primary procedure (key steps no details) that you will follow to clone the Cpeptide gene from the RNA above into the only vector you have, a pUC expression vector cut open using EcoR1 which has a 5recognition site and a 3HindIII in the MCS? 3) How can you guarantee a high expression of your protein in any expression vector?
You are given 3 tubes of RNA and asked by your supervisor to clone the gene encoding the proinsulin C-peptide that will be used later to produce insulin. The first tube contains purified mRNA isolated from pancreatic β-cells from a patient in a postabsorptive state. The second tube contains purified mRNA isolated from pancreatic α-cells from a patient during absorptive state. The third tube contains purified mRNA isolated from pancreatic β-cells from a patient during absorptive state. In not more than 2 pages (using 1.5 line space of Arial or Times New Roman fonts) provide answers for the following questions? 1) Which tube from the three is the most appropriate to use and why? 2) Describe the primary procedure (key steps no details) that you will follow to clone the Cpeptide gene from the RNA above into the only vector you have, a pUC expression vector cut open using EcoR1 which has a 5recognition site and a 3HindIII in the MCS? 3) How can you guarantee a high expression of your protein in any expression vector?
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