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Week10 8/15/10-8/21/10
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We continued working on the MAGE tqsA knockouts. We cloned CHS3 into a standard chlor biobricks backbone and began site directed mutagenesis (SDM) to remove the PstI site. | We continued working on the MAGE tqsA knockouts. We cloned CHS3 into a standard chlor biobricks backbone and began site directed mutagenesis (SDM) to remove the PstI site. | ||
Latest revision as of 23:06, 16 October 2010
Week10 Highlights
We continued working on the MAGE tqsA knockouts. We cloned CHS3 into a standard chlor biobricks backbone and began site directed mutagenesis (SDM) to remove the PstI site.
8/16/10
Cloned CHS3 into Chlor Biobricks plasmid --> will be used for Site Directed Mutagenesis
Cycles 2 and 3 of MAGE for tqsA Knockouts.
8/17/10
Site-Directed Mutagenesis to remove the PstI site in CHS3
Cycles 4-6 of MAGE for tqsA Knockouts.
8/18/10
Site-Directed Mutagenesis. Ran screening for 8/17, none were positive so repeated reaction with better protocol. (PCR only)
Cycle 7 of MAGE for tqsA Knockouts.
8/19/10
Site-Directed Mutagenesis. Transformed products from 8/18
Cycles 8 and 9 of MAGE for tqsA Knockouts.
8/20/10
Site-Directed Mutagenesis-Redo PCR. 10 colonies were screened, all negative (Two bands were seen when cut with PstI)
Cycles 10-12 of MAGE for tqsA Knockouts.
