User:Howard Boland/Notebook/Art from Synthetic Biology/2010/08/08

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Overnight Growth - of pUA66 and Ligation

Inoculate 8 tubes of 10ml LB broth w/Kanamycin using pUA66 Inoculate 2 tubes of 10ml LB broth w/Kanamycin using transformed colonies ligased with pUA66-XhoI-BamHI and katE-XhoI-BamHI. --Howard Boland 16:26, 8 August 2010 (EDT)


The colonies from this weekend growth show that the SAP (Shrimp Alkaline Phosphatase) treated vector did not work - only the standard NEB ligation. I am pretty sure that the ligation once digested will show my insert gone and the plasmid will remain either the pUA66 or a double ligation of the vector.

Too much time!

I am concerned about how much time it is taking to develop this project. I was expecting maximum one month. There issues might be raised as to why: 1) It is the problem of the project 2) It is a problem with technique or training 3) There are better ways of developing the project (synbio).

Should I ask to work in conjunction with my supervisor?


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