Team:TUDelft/1 September 2009

=1 September 2009=

Sriram
Today I continued with the final assembly of Riboregulator. Since Daniel is leaving I also continued his assembly of locks and keys. Today I sent samples to sequenceing (Medium RBS control, Plasmid 1 of negative cascade, Plasmid 2 of negative cascade and Weak RBS Control). I added both primers given by TIm Weenink.

Daniel
Miniprep of yesterday's cultures:

As noticed, 1C concentration is low, this is because I add neutralization buffer before lysis buffer. We will do it again tomorrow.Thanks to Esengul, we will have electrocompetent cells which produce LacI.

Today I leave the lab and I'll be helping in planning and experimental design.

Tim Weenink
Today I restricted p(LacI) as an upstream part and *T1 (homing endonuclease without promotor) as a downstream part. Additionally I ligated the restricted I-SceI PCR fragment. This fragment was PCR'd from the plasmid from leiden containing the conding sequence of ISCEI. It was purified and restricted using E and P. Using the standard protocol, it was ligated into a pSB1A3 vector.

Finally, Esengul streaked a plate of K12 strain, that allows IPTG induction (without being expressed constitutively). This strain wil be grown up and made electro competent.