Team:Panama/26 July 2010

July 26

We performed miniprep, for the E. coli transformed with the RBS, GFP and the promoter plasmid. We measured DNA concentration with a spectrometer. For this, we diluted the samples 1:100 in sterile water.

The measurements were performed with 260nm and 280nm wavelenght, we obtained:

[RBS]=55,7 ng/μl

[Promoter]= 71,8 ng/ul

For the analysis of the plasmid size, we prepared a 1% agarose gel.

Plasmids expected size:

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Agarose Gel 1%

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During the reporter extraction we faced some problems. We obtained a small amount of plasmid, probably because we cultured it for more than 16 hours and we obtained low plasmid yield. We couldn’t visualize the expected bands, probably because in the first miniprep, we did not obtain enough DNA concentration. For this reason, we inoculated more of the transformed E. coli colonies in LB broth for 16h at 37°C with vigorous shaking.