"
Team:Cornell/Week 19
From 2011.igem.org
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 |
September 25th - October 1st
Sunday, October 9
iGEM 2011 Regionals Jamboree: Americas!
Monday, October 10
- iGEM 2011 Regional Jamborees: Americas Award Ceremony
- Cornell received Gold Award
- Cornell advances to the iGEM 2011 World Jamborees!
- Outreach
- Attended meetings for both AlumniGEM and Community Bricks.
Tuesday, October 11
Wednesday, October 12
Lab work done by: Charlie Chung
- Picked Colonies: five (40mL LB + 40µL ampicillin) cultures shaking at 37°C
- - AviTagged GFP
- - AviTagged VioA from PCR deletion
- - AviTagged VioB from PCR deletion
- - AviTagged VioE from PCR deletion
Thursday, October 13
Lab work done by: Charlie Chung
- Freshly made 1000x ampicillin and 1M IPTG are in the DNA box of Olin 301 (-20°C fridge)
- 20mL from 40mL cultures of VioA, B, E were transferred to (1L LB + 1mL ampicillin) flask for a 1:50 dilution
- - Wanted OD to be between 0.05 and 0.08 after a 2-3hr incubation period, but accidentally let it grow over to 0.1xx
- - Regardless, induced with 1mL 1M IPTG
- - Shaking at room temp in Olin 301 for 20+ hours until 6pm on Friday (when we'll spin the cells down)
- two 10mL from 40mL culture of GFP were transferred to two (500mL LB + 500µL ampicillin) flasks for a 1:50 dilution
- - Wanted OD to be between 0.05 and 0.08 after a 2-3hr incubation period, but accidentally let it grow over to 0.1xx
- - Regardless, induced with 500µL 1M IPTG
- - Shaking at room temp in Olin 301 for 20+ hours until 6pm on Friday (when we'll spin the cells down)
- Took 500µL from remainder of the 40mL cultures and added 500µL of 30% glycerol to create freezer stocks of GFP and VioA, B, E
- - In -80°C freezer of Olin 303
- Miniprep purification of 5mL from remainder of the 40mL cultures. Eluted products are in -20°C fridge. To be submitted for sequencing
Friday, October 14
- Subculturing of Negative Control (empty pZE12 vector)
- Transfer two 10mL samples from 40mL culture of empty pZE12 vector to two (500mL LB + 500µL ampicillin) flasks for a 1:50 dilution
- Incubate overnight on 37°C shaker in Olin 304
- Protein Extraction of GFP and VioA, B, E
- Spin down 1L cultures into a pellet (10 min at 3200 rpm)
- Resuspend in 20mL autoclaved water
- Transfer to 50mL conical tube and spin for 10 min at 3200 rpm
- Store in -80°C freezer
- Western Blot Preparation
- Centrifuge 1.5mL of GFP and VioA, B, E cultures for 10 min at 14,000 rpm
- BugBuster will be used for protein extraction
