Alice:LabNotes/2008-5-2
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PCR[edit]
reaction system x2 H2O 32.1ul 64.2ul 10x JumpStartTaq Buffer 5ul 10ul MgCl2(25mM) 3ul 6ul dNTP(10mM) 2ul 4ul AmpF6.2/R6.2 (10uM) 2ul 4ul 50x SYBG I 0.4ul 0.8ul JumpStartTaq 0.5ul 1ul Products of Circluarization 5ul 10ul Total 50ul 100ul
94C 3min -> 20cycles of (94C 45sec -> 57C 1min -> 72C 1min) -> 72C 3min -> 4C hold
File:ZhangLab 2 2008-05-02 18hr 51min---.jpeg
The right most lane is the PCR product from 57.5C, and the second to the right lane is the product from 60C. Obviously, 57.5c has much more intense band and better yield. Also, since the primer is the one with tail, so the expected result should be around 150bp, which is consistent with result obtained.