From 2013.igem.org

June 07th 2013

Ligation or transformation from 6/6 did not work, the plates did not grow.  Brandon transformed the remaining 3:1 and 5:1 ligation reaction again.  Approximately 20 ng/uL of DNA is present in the digestion reactions, but it would be good to check on a gel anyway.  Gel was set up with a few uL of both the digested Aer/EnvZ and the digested OmpR.  A round of cleaning and concentrating may be needed before using it in another ligation reaction.  The transformation was continued as planned.  

The gel did not show any bands for either digested segment.  

Brandon set up another digestion.  Aer/EnvZ is digested with both BglII and XhoI.  OmpR is digested with NdeI and BamHI.  The digestion ran for 2.75 hours at 37 C.

1 uL of each enzyme. (2 uL total)

1 uL Buffer (NEB Buffer 3 for both)

2 uL Template

5 uL sterile DI water

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