From 2013.igem.org

July 18th 2013

The confirmation PCRs did not work. Confirmation digestion was negative as well. EcoRI and BamH1 were used for digestion. I am going to re-PRC entirely new pieces of backbone, Aer, EnvZ, OmpR, and the Aer-EnvZ chimera.

Aer and OmpR PCRs were unsuccessful. I ran a gel on all old DNA samples and only the old 5/30 Aer was still good. I am extracting EnvZ and Aer from the gel.

OEPCR:

12.5ul Q5 mm

150ng each template

7ul H20

*pause*

Add 1.25ul each primer.

25ul total

I grew up more PATBS01 to extract tomorrow as backup.

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