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From 2010.igem.org

Module One 1:

PCR for Tet, Landing Pad added. Pyrobest Enzyme

Module I, Group 2c

We are informed that the volume of PCR system somehow has an effect on the result. Usually, 25ul or 20ul is a better option than 50ul system. To further modify the condition for PCR, we compared the result of 25ul and 50ul system.

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PCR system (pfu):

 H2O	                       13ul
 5×buffer	               5ul
 dNTP	                       2.5μl
 primer1	               1μl
 primer2	               0.5μl
 template	               1μl
 FastPFU	               2μl
 Total	                       25μl

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PCR system (pfu):

 H2O	                       26ul
 5×buffer	               10ul
 dNTP	                       5μl
 primer1	               2μl
 primer2	               1μl
 template	               2μl
 FastPFU	               4μl
 Total	                       50μl

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Result: No specific target bands are seen in the gel photograph.