Difference between revisions of "Team:UCLA/Notebook/Honeybee Silk/11 August 2015"
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=8/11= | =8/11= |
Latest revision as of 22:02, 24 August 2015
Ligation of psb1c3+T7 promoter and Silk+GFP
Component | Volume (out of 20uL) |
---|---|
T4 Ligase Reaction Buffer | 2 ul |
Backbone (psb1c3+t7 promoter) | 0.8uL (50 ng) |
Insert (silk + spyC) | 1.5 ul (146 ng) |
H20 | 14.7 ul |
Ligase | 1uL |
- Program =16C overnight, 10 minutes heat inactivation at 65C.
8/11
Protein Yield
- Yield from 8/7 silk purification
- BCA assay
- Based on the interpolated Absorbencies from the BCA assay the concentration of our 8/7 silk is approximately 0.78 mg/ml in around 12 ml of solution, giving a conservative 9 total mg of protein for a 300 ml culture.
- This is an improvement over the 7/29 purification where we got around 1.6 mg/ml in only 0.75 ml of solution from a 100 ml culture.