Week10 8/15/10-8/21/10

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Week10 Summary

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.


Cloned CHS3 into Chlor Biobricks plasmid --> will be used for Site Directed Mutagenesis

Cycles 2 and 3 of MAGE for tqsA Knockouts.


Site-Directed Mutagenesis to remove the PstI site in CHS3

Cycles 4-6 of MAGE for tqsA Knockouts.


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.


Site-Directed Mutagenesis. Transformed products from 8/18

Cycles 8 and 9 of MAGE for tqsA Knockouts.


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.