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Team:The University of Alberta/22 July 2008
From 2008.igem.org
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* Did minipreps on O/N's of I0500 (new) col 1 and col 2. They are kept in Miniprep box 2 in freezer. | * Did minipreps on O/N's of I0500 (new) col 1 and col 2. They are kept in Miniprep box 2 in freezer. | ||
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| + | David digested BisdA and BisdB for insertion into 2 different vectors each: | ||
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| + | I digested BisdA and BisdB with both (NgoMIV+Age1) and (XbaI+PstI) and concentrated all of the digests down to 30ul. | ||
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| + | I digested the vector containing the TetR binding site and the RBS with NgoMIV and treated the digest with Antartic Phosphatase. I also concentrated this digest down to 30ul. | ||
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| + | I digested J61003 with (SpeI+PstI) and treated the digest with Antartic Phosphatase. I also concentrated this digest down to 30ul. | ||
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| + | The concentrated digests have been kept at -20C overnight and should be gel purified tomorrow, with the BisdA/B genes being purified and inserted into the digested vectors. | ||
Latest revision as of 01:38, 23 July 2008
Mike added links to two protein complementation papers in the Project/Journals section of the Wiki. One of these uses a Vibrio transcription factor (ToxR) fragment and Vibrio promoter in E. coli, and the other uses B-galactosidase in E. coli. Can anyone figure out how to make the appropriate BioBricks to fuse these to the existing ER LBD BioBrick? We should be able to amplify the B-galactosidase fragments from E. coli, but we will need to synthesize the Vibrio fragments and promoter. We will also need to order a lacZ deletion strain of E. coli (listed in the paper) if we don't already have something like it.
Can anyone commit to designing these primers and synthetic constructs (ASAP?)
- Did minipreps on O/N's of I0500 (new) col 1 and col 2. They are kept in Miniprep box 2 in freezer.
David digested BisdA and BisdB for insertion into 2 different vectors each:
I digested BisdA and BisdB with both (NgoMIV+Age1) and (XbaI+PstI) and concentrated all of the digests down to 30ul.
I digested the vector containing the TetR binding site and the RBS with NgoMIV and treated the digest with Antartic Phosphatase. I also concentrated this digest down to 30ul.
I digested J61003 with (SpeI+PstI) and treated the digest with Antartic Phosphatase. I also concentrated this digest down to 30ul.
The concentrated digests have been kept at -20C overnight and should be gel purified tomorrow, with the BisdA/B genes being purified and inserted into the digested vectors.
