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Team:Nevada/Week 2
From 2012.igem.org
Protocols | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Week 11 | Week 12 | Week 13 | Week 14 | Week 15 | Week 16 | Week 17 | Week 18 | Week 19 | Final Weeks |
Contents |
May 28
- Michelle:
- Ligation of SBP + LRP transformed onto Kanamyacin (KAN) plates. Gel analysis of SBP.
- Justin and Dafne:
- Digest SBP and B12 again.
- Initial digests were unsuccessful
May 29
- Jeremiah and Chris:
- Made new CCMB80 buffer for use in purification and new competent cells in SOB. Re-ligated SBP-TBP as gel showed no results for first attempt.
- Justin and Dafne:
- Ligate B12 into SBP
- SBP was dephosphorylated first, using Antarctic phosphatase
- Michelle and Joe:
- Check transformation of SBP + LRP from KAN plates. Made more competent cells.
May 30
- Michelle and Joe:
- Test competent cells and prepared for transformation of RFP in the future.
- Jeremiah and Chris:
- Testing for contamination in digests
May 31
- Michelle and Joe:
- Miniprep and digest LRP with PstI and XbaI and SBP with SpeI and PstI. Ran gel of LRP and SBP digestions.
- Colony PCR followed by ligation of SBP (SpeI and PstI) with LRP (PstI and XbaI).
- Transformation of RFP into competent cells.
- Jeremiah and Chris:
- Colony PCR done of ligated and transformed colonies of SBP-TBP
- Justin and Dafne:
- Digest plasmids again. Initial digests were unsuccessful
June 1
- Michelle and Joe:
- Transformation of SBP (SpeI and PstI) and LRP (PstI and XbaI) ligation onto KAN plates.
- Jeremiah and Chris:
- Competent cells tested to ensure transformability – did not work.
- Transformed SBP-TBP into competent cells
