Lab Notebook

4 August 2009

Sriram

We found that we need more concentration of parts' DNA to continue our assemblies. We are bit skeptical how far we will be successful in the ligated parts transformation. Hence we decided to try both the heat shock transformation and electro transformation on the ligated parts. Today I cultured colonies from plates of all parts needed for tomorrow's miniprep.

Orr

Prepared some LB Agar medium, and continued with writing the code for the software constructing lock and key sequences for the cascade delay device.

Calin

Trimethoprim finally arrived. Made trimethoprim 4 x 1 mL eppendorfs stock solution of 10 mg/mL using DMSO solution.

Transformed the first two ligations (CA & CB).

Made two R751 plates with 2xTRI.

Made a 5 mL tube of pTet-GFP with AMP and another 5 mL tube of R751 with TRI.

Tim Weenink

none of yesterdays transformations worked. To find out why, a gel was prepared with the DNA fragments that were used. Also, a PCR reaction of the all the colonies from !D transformation was done. For this the primers I-SceI forward and VR were used. Thirdly, *S, *T, *E, *G, *P PCR products were purified. This was done because it was found that adding 10µl of PCR reaction mix directly to the restriction mix will cuase star activity.

Results of the gel are on next days page.

results of the purification: