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Team:Cornell/Week 6
From 2011.igem.org
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#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. | #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 | #Animations and website are continuing | ||
| + | *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. | ||
| + | *Didi will take PCR product of RFP and see if amplification worked via gel electrophoresis. If yes, then she will gel extract. Probably leave the gel purification steps for us? NOTE: P.O.S. 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 Bootcamp Packet | ||
==Saturday== | ==Saturday== | ||
__NOTOC__ | __NOTOC__ | ||
Revision as of 03:51, 18 July 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 |
June 10th - June 16th
Sunday
- Worked on Website. Preliminary design of banner, safety content, and temporary lab notebook spreadsheet.
- Ligation product was transformed in electrocompetent cells, culture plated on Amp
- Vio-Operon plasmid was transformed and plated on Kanamyacin
- RFP PCR done overnight
Monday
Tuesday
Wednesday
Thursday
- 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
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
- 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.
- Didi will take PCR product of RFP and see if amplification worked via gel electrophoresis. If yes, then she will gel extract. Probably leave the gel purification steps for us? NOTE: P.O.S. 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 Bootcamp Packet
