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Team:KULeuven/16 July 2008
From 2008.igem.org
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Lab Work
Wet Lab
Yesterday's transformation gave no colonies. Today, we performed the same transformation ([http://partsregistry.org/Part:BBa_M30109 M30109]) in competent TOP10 cells using the BioBrick protocol (and not the CMPG protocol). An empty pUC plasmid was used as control. Because the transformation didn't succeed yesterday, we are a bit concerned about the use of M30109. We found that no project before could ever succesfully use this part.
[http://partsregistry.org/Part:BBa_B0034 B0034] was punched out and transformed into competent DH5alpha cells using the CMPG protocol, also with empty pUC control.
Buffer CCMB80, SOB agar and SOB broth were made. Incompetent cells were streaked out on a SOB agar and the first step in making them competent was conducted, following the Registry's protocol.
Dry Lab
General
We did some research concerning the UmuD tag we need for the T7 polymerase. UmuD should reduce the half-life of T7 polymerase to approximately 9 minutes. The sequence in FASTA-format and more information about the UmuD tag, is listed under 'Literature'.
Modeling
Parameters... Making very significant progress.
