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Team:Cornell/Week 7
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{{:Team:Cornell/Templates/Menu}} | {{:Team:Cornell/Templates/Menu}} | ||
| - | {{:Team:Cornell/Templates/Notebook}} | + | {{:Team:Cornell/Templates/hideHeader}} |
| + | {{:Team:Cornell/Templates/MediaMenu}} | ||
| + | {{:Team:Cornell/Templates/Notebook}} | ||
<br> | <br> | ||
<html> | <html> | ||
| - | <font size="5"> <i> July 17th - July 23rd <i></font> | + | <font size="5"> <i> July 17th - July 23rd </i></font> |
</html><br><br> | </html><br><br> | ||
| - | ==Sunday== | + | ==Sunday, July 17== |
| - | * | + | Lab work done by: Alyssa Henning, Bill Jo |
| - | *Ran a gel on the RFP that was | + | :*Miniprepped 1 sample of GFP + AviTag (the second sample got messed up) |
| - | *Sean will | + | :*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== | + | ==Monday, July 18== |
| - | *Gel purified 2 RFP PCR samples from Friday | + | Lab work done by: Charlie Chung, Youjin Cho |
| - | * | + | :*Gel purified 2 RFP PCR samples from Friday |
| + | :*Miniprep 2 samples of GFP + AviTag | ||
| - | ==Tuesday== | + | ==Tuesday, July 19== |
| + | Lab work done by: James Mathew | ||
| + | :*Submitted GFP + AviTag for synthesis | ||
| + | :*Submitted pZE12 backbone for subcloning of light sensor | ||
| - | ==Wednesday== | + | ==Wednesday, July 20== |
| + | Lab work done by: James Mathew | ||
| + | *'''Set up ligation reaction of pZE-12 backbone with the primer dimer insert''' | ||
| + | ::1. <u>Digestion of Backbone</u> | ||
| + | :::- 26µl Backbone plasmid = 2µg DNA | ||
| + | :::- 5µl NEBuffer 4 (optimal for SphI-HF and ClaI-HF) | ||
| + | :::- 1µl SphI & 1µl ClaI | ||
| + | :::- 16.5µl H2O | ||
| + | ::::*left in water bath for one hour | ||
| + | :::- 1µl CIAP added to digestion reaction | ||
| + | ::::*left in water bath for 30 minutes | ||
| + | ::2. <u>Ran Digestion Product through gel for 30 minutes at 120V</u> | ||
| + | ::::Lane 1: 2µl 1kb Ladder + 2µl 6x loading dye + 8µl H2O | ||
| + | ::::Lane 2: 25µl digestion reaction + 5µl 6x loading dye | ||
| + | ::::Lane 3: 25µl digestion reaction + 5µl 6x loading dye | ||
| + | ::3. <u>Gel Purification using Qiagen Kit</u> | ||
| + | ::4. <u>NanoDrop of Samples</u> | ||
| + | ::5. <u>Ligation Reaction</u> | ||
| + | :*Ligation done overnight for transformation on 7/21/11 | ||
| - | ==Thursday== | + | ==Thursday, July 21== |
| + | Lab work done by: James Mathew | ||
| + | *Miniprepped Vio operon (Cambridge 2009) | ||
| + | Lab work done by: Youjin Cho, Charlie Chung, Alyssa Henning | ||
| + | *Reconstituted VioA, VioB, and VioE forward and reverse primers | ||
| + | *Set up PCR for VioA, VioB, and VioE | ||
| + | *Aborted transformation of pZE12 plasmid + AviTag primer dimer because reverse primers for AviTag were incorrect | ||
| + | *Redesigned reverse primers for AviTag to order on Friday | ||
| - | ==Friday== | + | ==Friday, July 22== |
| - | + | [[File:Cornell11 VioA,B,E PCR 7-22.jpg|thumb|Gel electrophoresis]] | |
| - | + | <br> | |
| + | Lab work done by: James Mathew, Claire Paduano | ||
| + | <br><br> | ||
| + | *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 | ||
Latest revision as of 17:23, 18 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 for synthesis
- Submitted pZE12 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
- - 26µl Backbone plasmid = 2µg DNA
- - 5µl NEBuffer 4 (optimal for SphI-HF and ClaI-HF)
- - 1µl SphI & 1µl ClaI
- - 16.5µl H2O
- left in water bath for one hour
- - 1µl CIAP added to digestion reaction
- left in water bath for 30 minutes
- 2. Ran Digestion Product through gel for 30 minutes at 120V
- Lane 1: 2µl 1kb Ladder + 2µl 6x loading dye + 8µl H2O
- Lane 2: 25µl digestion reaction + 5µl 6x loading dye
- Lane 3: 25µl digestion reaction + 5µl 6x loading dye
- 3. Gel Purification using Qiagen Kit
- 4. NanoDrop of Samples
- 5. Ligation Reaction
- Ligation done overnight for transformation on 7/21/11
- 1. Digestion of Backbone
Thursday, July 21
Lab work done by: James Mathew
- Miniprepped Vio operon (Cambridge 2009)
Lab work done by: Youjin Cho, Charlie Chung, Alyssa Henning
- Reconstituted VioA, VioB, and VioE forward and reverse primers
- Set up PCR for VioA, VioB, and VioE
- Aborted transformation of pZE12 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
