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Team:Nevada/Week 12
From 2012.igem.org
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==August 6== | ==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== | ==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 | ||
| + | ::: | ||
==August 8== | ==August 8== | ||
Revision as of 19:41, 25 September 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 | Week 20 | Week 21
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
