Lab Notebook

8 September 2009

Sriram

Today I got in my positive control both GFP glowing and non-GFP glowing K12 strain colonies which is not good. Hence I cultured them today so that I can miniprep and analyse tomorrow. I got good colonies for the final assemblies for locks and keys final assemblies. They also must be checked in gel tomorrow hence I cultured them too. Also we think its good time to assemble a LacI generator which has 2 assembly steps. Hence I prepared it too. I also cultured some of the electrotransformed sample to the culture so that I can get the LacI generator faster.

Tim Weenink

Today: Plasmid isolation: out of 10 !SIII inoculated cultures, only 3,5 and 6 showed proper growth. The others showed very low growth and will be incubated for more time. Growth/low growth is not related to colony size, which seemed to have two growing modes: big colonies and small colonies.

Did miniprep and made glycerol stock and did E-P restriction of the following:

Also did E-P restriction of the pSB4C5 backbone. This was all gelled:

Finally I did a ligation of pLacI (upstream), *T1 (BBa_K142202: I-SceI homing endonuclease with RBS & double terminator) (downstream) and pSB4C5 as a backbone.