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From 2010.igem.org

Module I, group 2(b)

PCR and double digestion to detect.

Double digestion system:

 H2O	        8μl
 buffer tango	4μl
 plasmid	5μl
 EcoRI	        1μl
 BamHI	        2μl
 Total	        20μl

PCR system (SuperMix):

 H2O	        8.5μl
 primer1	0.5μl
 primer2	0.5μl
 template	0.5μl
 SuperMix	10μl
 Total	        20μl

result

The result is quite strange. We’ll buy a pair of pUC19 universal primers to run the PCR. Also, we doubt that the PEM plasmids are not pure. So we transfer the plasmids PEM again to purify.