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Exeter/29 August 2013
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Revision as of 11:06, 2 September 2013 by JoshMiddleton (Talk | contribs)
Minipreps
We miniprepped:
- 1 + 4
- K592011 B
- K864404
- Cph8 #3
- 2 + 4
- K592022 B
- ENvZ no antibiotic
- ENvZ
- Cph8 #5
The liquid cultures of K592011 A and K592022 A did not grow.
NanoDrop
We collected NanoDrop data of previously miniprepped, defrosted DNA. The NanoDrop machine was zero-ed using the elution buffer from the gel extraction kit used.
- OmpF A - 20.1 ng/ul
- OmpF B - 9.3ng/ul
- B0034 - 26.7ng/ul
- RFP - 198ng/ul
The concentration for OmpF B was too low, so we continued on to the digestion with only OmpF A. The concentration for RFP was too high, so we diluted it.
- RFP (dilution) - 50.4ng/ul
Digestion
| Part | OmpF (Part A) | B0034 (Part B) | AMP Plasmid | RFP (Control) |
|---|---|---|---|---|
| DNA | 12.4 ul | 9.4 ul | 10.0 ul | 5.0 ul |
| Non-nuclease Water | 3.6 ul | 6.6 ul | 5.5 ul | 11.0 ul |
| NEB Buffer | 2.5 ul | 2.5 ul | 2.5 ul | 2.5 ul |
| BSA | 0.5 ul | 0.5 ul | 0.5 ul | 0.5 ul |
| EcoR1 | 0.5 ul | - | 0.5 ul | 0.5 ul |
| Spe1 | 0.5 | - | - | - |
| Xbal1 | - | 0.5 ul | - | - |
| Pst1 | - | 0.5 ul | 0.5 ul | 0.5 ul |
| Dpn1 | - | - | 0.5 ul | - |
