Difference between revisions of "Team:UCLA/Notebook/Honeybee Silk/10 July 2015"
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Revision as of 02:19, 11 July 2015
Cloning Honeybee Silk into Pet 24a
- Sequencing results checked out for miniprepped pet 24a plasmid.
- Double digested PET 24a with BamH1 and Nde1 restriction enzymes
- Ran on gel, excised, and purified
- Ran ligation reaction with double digested pet 24a and honeybee sequenced
| Component 3:1 insert:vector) | Volume (out of 20uL) |
|---|---|
| T4 Ligase Buffer | 2 ul |
| Pet 24a backbone (23.6 ng/ul) | 2.54 ul |
| SIlk insert 28.4 ng/ul | 1.12 |
| T4 Ligase | 1 ul |
| Water | 13.34 |
| Component 5:1 insert to vector | Volume (out of 20uL) |
|---|---|
| T4 Ligase Buffer | 2 ul |
| Pet 24a backbone (23.6 ng/ul) | 2.54 ul |
| SIlk insert 28.4 ng/ul | 1.9 |
| T4 Ligase | 1 ul |
| Water | 12.56 |
Running ligation reaction overnight 16 C starting at 6:45 pm 7/10