Question

Assume you will run a PCR on a 1Kb DNA fragment. Assume your DNA template's upper strand is the 5' to 3' strand. The primers you will use are P1 and P2. The 5’ end of P1 recognizes sequence # 190. The 5’ end of P2 recognizes sequence # 350. The PCR conditions are 95oC for 1 minute, 55oC 30 seconds and 72oC for 30 seconds for 30 cycles. (a) indicate the size of your PCR product; (b) which primer, P1 or P2, binds to the upper DNA strand of your template?; (c) explain the need to use 95oC and the reagents involved (if any) in the reaction at this temperature; (d) explain the need to use 55oC and the reagents involved (if any) in the reaction at this temperature; (e) explain the need to use 72oC and the reagents involved (if any) in the reaction at this temperature.