Team:UCLA/Notebook/Protein Cages/18 August 2015
Phillip's notes:
Introduction: The gBlocks for the new mutants will be amplified.
Procedure: Nithin ran the PCR to amplify. See his section for details.
Running on the gel:
The two 50uL reactions were pooled and 20uL from each was mixed with 5uL of 5x loading dye. The gel was loaded and ran for 20 minutes at 100V.
The results of the gel indicated successful amplification of all mutants. From left to right: Ladder, Mutant 1, Mutant 2, Mutant 4, Mutant 13. Media:2015-08-18_PCR_amplification_M1,_M2,_M4,_M13.JPG
PCR clean up was done on the remaining sample from the reaction.
The nanodrop was used to measure protein concentration, 260/230, and 260/280.
Mutant 1: 134.04ng/uL, 1.87, 1.88.
Mutant 2: 145.36ng/uL, 2.00, 1.83.
Mutant 4: 123.45ng/uL, 1.67, 1.91.
Mutant 13: 124.58ng/uL, 1.97, 1.84.
Conclusion: Digestion, and ligation should be done tomorrow.