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Team:Nevada/Week 12
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 |
Contents |
August 6
- Joe: colony check SBP-RFP + terminator
- Michelle:
- Check transformation of SBP (SpeI and PstI)-LRP (PstI and XbaI) digested with XbaI
- and PstI and CP-EP* L-arabinose (SpeI and NSI).
Miniprep SBP (SpeI and PstI)-LRP (PstI and XbaI) and digest with SpeI and PstI.
- Jeremiah & Chris:
- Colony PCR done to check if the terminator added successfully (gel #133)
- Grow colony 12
- Digest TBP+ again incase colony 12 fails
August 7
- Joe: miniprep SBP-RFP-Terminator
- digest by EcoRI/PstI to check, digest by EcoRI/ XbaI for ligation
- Gel 139- both digestions good
- Ligate SBP-RFP-Terminatior and promoter
- Transform, plate on Kana
- Michelle:
- Run gel to check digest SBP (SpeI and PstI)-LRP (PstI and XbaI) with SpeI and PstI,
- heat inactivate, and dephosphorylate digest.
- Ligate SBP (SpeI and PstI)-LRP (PstI and XbaI) with SpeI and PstI digest with
- Terminator (PstI and XbaI) digest followed by transformation into TOP 10 competent cells onto KAN plates.
- Justin and Dafne
- Clean corn starch 3x
- Place 200 ul of refolded protein in 1mg/ml solution of corn starch in 1.5 ml epindorf tube
