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Team:Brown/Notebook/July14
From 2010.igem.org
Wednesday, July 14 2010
Ran the PCR product on a 1% agarose gel
- Lane 1 – 1kb+ ladder
- Lane 2 – iGEM pGEM
- Lane 3 – Gary pGEM
Result: blank gel; saw ladder (not worth imaging) but neither iGEM pGEM nor Gary pGEM; implies that our suspicions were correct and we had no insert in our blue-white selection.
Now we need to decide on which step to start over.
Transformation of BBa_J06702
- 15:05 – retrieved 2 aliquots of Bl21s and 2 aliquots of XL1-Blues.
- Followed ‘quick competent cell procedure’ (altered) from this notebook to make overnight culture tube of Xl1-Blues from last night. Did not see any pellets during the centrifuge steps, even though the OD went up every 20 minutes until ~0.3 OD at 600 nm, so we carried on with the procedure.
- Followed altered transformation protocol for both fresh cells and frozen stock and placed plates in the incubator.
- pPTPi transformants from 7/13 grew in a lawn. Hypothesized problem: Zach’s tet was shot. Using our tetracycline, we repoured 23 LB plates at 10 µg/mL
- Transformed pPTPi and RFP controls as we did on 7/13.
- Incubate overnight starting at 8:00 PM.
