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==Experiment: CES22K probe preparation using Jumpstart Taq== reaction system 1: x48 H2O 80.5ul 3864ul 10x Stoffel buffer 10ul 480ul MgCl2(25mM) 6ul 288ul dNTP(10mM each, NEB) 2ul 96ul primer mix (Ap6) 0.4ul 19.2ul 50x SYBG I 0.4ul 19.2ul template(CES22K #4) 0.1ul 4.8ul Jumpstart Taq 0.6ul 28.8ul Reaction system 2: H2O 80.7ul 3873.6ul 10x Stoffel buffer 10ul 480ul MgCl2(25mM) 6ul 288ul dNTP(10mM each, NEB) 2ul 96ul primer mix (Ribo V6 100uM) 0.2ul 9.6ul 50x SYBG I 0.4ul 19.2ul template(CES22K #4) 0.1ul 4.8ul Jumpstart Taq 0.6ul 28.8ul 94c 2min -> 94c 30sec -> 60C 2min -> 72C 1min-> Plate read -> (94c 30sec -> 60C 2min -> 72C 1min)x 20 cycles -> 72C 5min -> 15C hold PCR curve: [[Image:7-31-08.jpg]] The reaction system 2 is unsuccessful because the forward primer was not added.
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