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.