Berk2010-Christoph: Difference between revisions
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Ligated isolated Bjh1882 and Bca1091 into above vectors. Transformed into Lefty strains. <br> | Ligated isolated Bjh1882 and Bca1091 into above vectors. Transformed into Lefty strains. <br> | ||
Need to ligate Bjh2245 (Missing pMLL6, need to digest). <br> | Need to ligate Bjh2245 (Missing pMLL6, need to digest). <br> | ||
TO DO: Digest pMLL6 with Eco/Bam and gel purify. Then ligate with Bjh2245. <br> | TO DO: | ||
*Digest pMLL6 with Eco/Bam and gel purify. Then ligate with Bjh2245. <br> | |||
*Check transformed Bjh1882 and Bca1091. | *Check transformed Bjh1882 and Bca1091. | ||
*Transform ligated Bjh2245+pMLL6 into Righty strain | *Transform ligated Bjh2245+pMLL6 into Righty strain | ||
Revision as of 17:22, 7 June 2010
Christoph Neyer 15:54, 7 June 2010 (EDT)
Eco/Bam digest of parts: Bjh2245 (LifeACT), Bjh1882 (RFP), and BCA1091 (Ptet).
Gel purified Bjh1882 and Bjh2245.
Small fragment zymo cleanup of BCA1091.
Christoph Neyer 16:35, 7 June 2010 (EDT)
Couldn't see fragments. Turns out Bjh2245 is 97bp and Bjh1882 is ~680bps.
Re-cut Bjh2245 and Bjh1882.
Small fragment zymo cleanup of Bjh2245 digest.
Gel purified Bjh1882.
Christoph Neyer 19:30, 7 June 2010 (EDT)
Bjh1882 Vectors: AC
Bjh2245 Vectors: KA
Bca1091 Vectors: CK
pMLL4-AC
pMLL5-CK
pMLL6-KA
pMLL7-AK
pMLL8-KC
pMLL9-CA
Bjh1882+pMLL4-AC = Lefty
Bjh2245+pMLL6-KA = Righty
Bca1091+pMLL5-CK = Lefty
Ligated isolated Bjh1882 and Bca1091 into above vectors. Transformed into Lefty strains.
Need to ligate Bjh2245 (Missing pMLL6, need to digest).
TO DO:
- Digest pMLL6 with Eco/Bam and gel purify. Then ligate with Bjh2245.
- Check transformed Bjh1882 and Bca1091.
- Transform ligated Bjh2245+pMLL6 into Righty strain
