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Team:Cornell/Week 7
From 2011.igem.org
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==Monday, July 18== | ==Monday, July 18== | ||
| - | Lab work done by: Charlie Chung | + | Lab work done by: Charlie Chung, Youjin Cho |
| - | *Gel purified 2 RFP PCR samples from Friday | + | :*Gel purified 2 RFP PCR samples from Friday |
| - | * | + | :*Miniprep 2 samples of GFP + AviTag |
==Tuesday, July 19== | ==Tuesday, July 19== | ||
Revision as of 01:52, 16 September 2011
Week 1 | Week 2 - 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 |
July 17th - July 23rd
Sunday, July 17
Lab work done by: Alyssa Henning, Bill Jo
- Miniprepped 1 sample of GFP + AviTag (the second sample got messed up)
- Ran a gel on the RFP that was amplified via PCR on Saturday
- Gel purification and extraction
- Sean will inoculate 6 tubes of GFP-containing bacteria -- 1 tube for us, 5 tubes for CURIE (Cornell iGEM's outreach program)
Monday, July 18
Lab work done by: Charlie Chung, Youjin Cho
- Gel purified 2 RFP PCR samples from Friday
- Miniprep 2 samples of GFP + AviTag
Tuesday, July 19
Lab Work Done By: James Mathew
- Submitted GFP+avitag genes for synthesis
- Submitted pZE-12 backbone for subcloning of light sensor
Wednesday, July 20
Lab work done by: James Mathew
- Set up ligation reaction of pZE-12 backbone with the primer dimer insert.
- 1. Digestion of Backbone
- - 26 ul Backbone plasmid = 2 ug DNA
- - 5 ul Buffer #4 (optimal for SphI-HF and ClaI-HF)
- - 1 ul SphI & 1 ul ClaI
- - 16.5 ul H20
- left in water bath for one hour
- - 1 ul CIAP added to digestion reaction
- left in water bath for 30 minutes
- 1. Digestion of Backbone
- 2. Ran Digestion Product through gel for 30 minutes at 120 V.
- Lane 1: 2 ul 1kb Ladder + 2 ul 6X Dye + 8 ul H20
- Lane 2: 25 ul digestion reaction + 5 ul 6X Dye
- Lane 3: 25 ul digestion reaction + 5 ul 6X Dye
- 2. Ran Digestion Product through gel for 30 minutes at 120 V.
- 3. Gel Purification using Qiagen Kit
- 4. NanoDrop of Samples
- Concentration:
- Label:
- 4. NanoDrop of Samples
- 5. Ligation Reaction
- Label:
- 5. Ligation Reaction
- Ligation done overnight for transformation on 7/21/11
Thursday, July 21
Lab work done by: James Mathew
- miniprepped Vio operon (Cambridge 2009)
Lab work done by: Youjin Cho, Charlie Chung, and Alyssa Henning
- Reconstituted VioA, VioB, and VioE forward and reverse primers
- Set up PCR for VioA, VioB, and VioE
- Aborted transformation of pZE-12 plasmid + avitag primer dimer because reverse primers for avitag were incorrect.
- Redesigned reverse primers for avitag to order on Friday
Friday, July 22
Lab work done by: James Mathew & Claire Paduano
- PCR gel purification of VioA, VioB, and VioE
Saturday, July 23
Lab work done by: James Mathew & Claire Paduano
- Digestion of VioA, VioB, VioE, GFP, and RFP in preparation for ligation with corrected backbone and avitag primer dimer
