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Team:Nevada/Week 7
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 |
July 2
- Jasmine, Joe, and Michelle:
- Amplified CP using PCR (adjusted amount of template)
- Digested EP* with SpeI and NsiI
- Ligated RFP* and EP*
- Jeremiah & Chris:
- Colony PCR of SBP-TBP in ExV (colonies 4 & 5 may be positive)
- Cultured LRP
- Digested TBP++ with SpeI and PstI
July 3
- Jasmine, Joe, and Michelle:
- Digested CP with EcoRI and SpeI
- Digested EP* with EcoRI and SpeI
- Amplified more CP using PCR
- Transformed RFP*-EP* ligation and plated onto Amp plate
- Justin and Dafne
- Miniprep SBP-B12(in expression plasmid) colonies
- Digest with Spe I to ensure the correct plasmid size
- Resulting electrophoresis analysis indicated successful creation of SBP-B12 in expression plasmid construct
July 4
- Jeremiah & Chris:
- Both standard and Phusion ligation of SBP-TBPa EXVTransform LRP-TBP++
July 5
- Jasmine and Joe:
- Checked 8 RFP*-EP* colonies with colony PCR
- Cultured colonies 7 and 8
- Amplified more RFP* using PCR
- Digested CP with EcoRI and SpeI (no purification after PCR)
- Gel purified EP*digested with EcoRI and SpeI
- Jeremiah & Chris
- Gel purified EP*digested with EcoRI and SpeI
- aDigest SBP-TBP for expression (again)with XbaI and PstI
July 6
- Jasmine and Joe:
- Dephosphorylated EP*
- Ligated CP and EP*
- Transformed into competent cells and plated onto Amp plate
