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Team:Nevada/Week 7
From 2012.igem.org
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[[Team:Nevada/Week 18| Week 18]] | | [[Team:Nevada/Week 18| Week 18]] | | ||
[[Team:Nevada/Week 19| Week 19]] | | [[Team:Nevada/Week 19| Week 19]] | | ||
| - | [[Team:Nevada/ | + | [[Team:Nevada/Final Weeks| Final Weeks]] | |
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==July 3== | ==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== | ==July 4== | ||
| + | :Jeremiah & Chris: | ||
| + | :::Both standard and Phusion ligation of SBP-TBPa EXVTransform LRP-TBP++ | ||
| + | |||
==July 5== | ==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== | ==July 6== | ||
| + | :Jasmine and Joe: | ||
| + | :::Dephosphorylated EP* | ||
| + | :::Ligated CP and EP* | ||
| + | :::Transformed into competent cells and plated onto Amp plate | ||
Latest revision as of 01:34, 27 October 2012
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 |
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
