Team:Tsinghua/Notebook/10 August 2010

From 2010.igem.org

Module I, DT and Fan's part:

To ensure the result again, we decide to run a PCR with each gene’s own primers.

PCR system (SuperMix):

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

And just before receiving the result, we also go on the experiments repeat the preview steps in case that we get the all negative result. One is to select another 12 colonies to amplify; another is to digest the eGFP, chlr and pUC19 again with the same system shown before.

In the afternoon, when the PCR has finished, run a 1.2% gel. The result is shown below.

Since the all 12 are all positive clones, stop the digestion and amplification step and go on the experiments. In consideration of the concentration of the plasmids and also the purity, we use P+E① and P+C③ undergo the follow steps.