Team:UCLA/Notebook/Recombinant Expression
Recombinant Silk Functionalization
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mm/dd: Sample Entry
Today we began cloning our GFP.
- PCR'd off template
- Ran gel
- Restriction digest
- Ligated into backbone
For our ligation, we made the following modifications:
- Tried it with newly bought ligase
- Left reaction overnight instead of 2 hrs
- Vector to insert ratio was 1:5 instead of 1:3
PCR Reaction:
Component | Volume |
5X Q5 Reaction Buffer | 5 |
10 mM dNTPs | 0.5 |
10 uM Forward (primer 3/7) | 1.25 |
10 uM Reverse (primer 8) | 1.25 |
Template (diluted to 1ng/uL) | 0.5 |
Q5 High Fidelity DNA Polymerase | 0.25 |
Nuclease Free Water | 16.25 |
Gel: Lot of bands, all at correct sizes
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