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Team:Cornell/Week 6
From 2011.igem.org
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==Sunday, July 10== | ==Sunday, July 10== | ||
Lab work done by: James Mathew, Charlie Chung | Lab work done by: James Mathew, Charlie Chung | ||
| - | + | :*Worked on website. Preliminary design of banner, safety content, and temporary lab notebook spreadsheet. | |
| - | + | :*Ligation product was transformed in electrocompetent cells | |
| - | + | :*Culture plated on agar plate treated with ampicillin | |
| - | + | :*Vio-Operon plasmid was transformed and plated on agar treated with kanamycin | |
| - | + | :*Overnight PCR reaction for RFP | |
==Monday, July 11== | ==Monday, July 11== | ||
Revision as of 01:15, 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 10th - July 16th
Sunday, July 10
Lab work done by: James Mathew, Charlie Chung
- Worked on website. Preliminary design of banner, safety content, and temporary lab notebook spreadsheet.
- Ligation product was transformed in electrocompetent cells
- Culture plated on agar plate treated with ampicillin
- Vio-Operon plasmid was transformed and plated on agar treated with kanamycin
- Overnight PCR reaction for RFP
Monday, July 11
Tuesday, July 12
- Finished final design of team logo
Wednesday, July 13
- "Website Team" meeting to continue design and construction of website
- - Designed and constructed final banner design
Thursday
Lab work done by: James Mathew & Charlie Chung
- Sequencing revealed no GFP. Retry the 3-piece ligation of GFP + Annealed Primer Pair A (half of biotin tag) + Annealed Primer Pair B (other half of biotin tag & iGEM suffix).
- PCR to amplify vector backbone containing RFP.
- Met Prof. Lucks -- prospective iGEM adviser and coolest man alive
Friday
- Banner was updated to include flash animation of moving gears.
Team Meeting
- Ligation didn't work, need to retry it over the weekend (transform and plate today, innoculate and colony PCR saturday, miniprep and prepare for sequencing on sunday).
- Prepare more culture of backbone plasmid (innoculate today, miniprep on saturday)
- We want to order primers today! for other methods of ligation, with longer primers, and two step PCR with short primers, in case our current one doesn't work. Also going to order primers for our pathway enzymes. The primers should come Wednesday. If ligation this weekend fails, we can try these other PCR methods.
- Animations and website are continuing
Lab work done by: James Mathew, Charlie Chung & Youjin Cho
- Running low on pZE vector backbone, so borrowed two culture tubes from Sean. Miniprep to purify out the plasmid. Followed standard Qiagen Miniprep protocol for microcentrifuge.
- Verify if PCR amplification of RFP worked using gel electrophoresis. If yes, then proceed to gel extraction and purification. NOTE: thermocycler did not keep our rxn tube @ 4 deg C
- Electroporation method of transforming our 3-piece ligation product and control into DH5α (competent E. coli = made porous for uptake of outside DNA). Plated transformed bacteria onto LB + Ampicillin. Followed protocol as written in the "Multimedia" section.
Saturday
Lab work done by: Youjin Cho & Nicholas Kramer
- Looked at the plates for ligation. Ligation worked!! Sean will pick 2 colonies off the plate in the evening and innoculate overnight.
- PCR to amplify vector backbone containing RFP.
- Melting temperature of 57C leads to non-specific binding. We ran 2 PCR reactions with melting temperatures 59C and 61C.
