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==comparison of PCR result using different Ribo concentration== reaction system 1: x48 H2O 80.1ul 3844.8ul 10x Stoffel buffer 10ul 480ul MgCl2(25mM) 6ul 288ul dNTP(10mM each, NEB) 2ul 96ul Primer (APV6) 0.4ul 19.2ul primer (Ribo V6 100uM) 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 10x Stoffel buffer 10ul MgCl2(25mM) 6ul dNTP(10mM each, NEB) 2ul primer (Ribo V6 100uM) 0.2ul primer (APV6) 0.2ul 50x SYBG I 0.4ul template(CES22K #4) 0.1ul Jumpstart Taq 0.6ul 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: TBE gel Result: [[Image:8-13-08.jpg]] [[Image:2008-08-13 22K with ribo V6.jpg]] Note:error on above image, the ladder should be 25bp instead of 10bp.
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