IGEM:University of Chicago/2009/Notebook/PNP Biosensor/2009/08/12

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August 12th, 2009

Something went wrong with my digestion yesterday, so I restarted it at 11:00 p.m. At 3:30 pm I added 2.5 uL SAP and 2.5 uL SAP buffer (found in Kron lab Freezer) to the template only to improve ligation efficiency.

My PCR Reactions weren't working because I was using the Kan-R primer, but my Longtine insert had the HIS marker. redoing this today with something Satoe gave me for reverse primer.

Make 1kb (f) DNA Ladder from Schonbam's sotck:

1. 2.5 uL 1 kb ladder

2. 87.5 uL milliQ

3. 10 ul B or H buffer (I used B)

4. 10 uL DNA loading Dye

Fragment sizes go from 12 kb to 0.16 (or 16???) kb or so.

End of the Day