You must design primers to amplify IDH1 using the polymerase chain reaction. The PCR experiment requires a forward primer, which anneals to the template (antisense) strand so that its 5' end is at the beginning of the IDH1 gene. You also require a reverse primer, which anneals to the other strand of the DNA (the coding, or sense strand). The reverse primer will have its 5' end at the end of the gene. As mentioned in the previous question, the primers should incorporate a restriction site to facilitate cloning into a plasmid vector. However, for the purpose of this question, do NOT include restriction sites into your primers. (Since the portions of the primers containing the restriction sites do not hybridize with the DNA being amplified their sequences do not affect the melting temperature of the primer-template complex.) The primers should be 15-25 bases long in order to ensure specificity in the annealing stage of the PCR experiment. Design your primers so that each will have a melting temperature of 62 °C. Calculate the melting temperature using the formula: Tm = 2° x (A + T) + 4° x (G + C)
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You must design primers to amplify IDH1 using the polymerase chain reaction. The PCR experiment requ...
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