Team:Cornell/Week 19

From 2011.igem.org

(Difference between revisions)
(Thursday, October 13)
(Monday, October 10)
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==Monday, October 10==
==Monday, October 10==
:'''iGEM 2011 Regionals Jamboree: Americas Award Ceremony'''
:'''iGEM 2011 Regionals Jamboree: Americas Award Ceremony'''
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::*Cornell received Gold Award
+
::*Cornell brings home the Gold
::*Cornell advances to the iGEM 2011 World Jamborees!
::*Cornell advances to the iGEM 2011 World Jamborees!

Revision as of 10:21, 15 October 2011

Results | Protocol | Notebook | Parts Submitted

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 Regionals Jamboree: Americas Award Ceremony
  • Cornell brings home the Gold
  • 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: four (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

Picking Colonies
  • (40mL LB + 40µL ampicillin) culture of empty pZE12 vector as a negative control shaking at 37°C
Miscellaneous
  • Freshly made 1000x ampicillin and 1M IPTG are in the DNA cryobox of Olin 301 (-20°C fridge)
Subculturing of AviTagged Vio Enzymes
  • 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 0.05-0.08 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)
Subculturing of AviTagged GFP
  • 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 0.05-0.08 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)
Preparing Freezer Stocks
  • 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
Preparation for Sequencing Confirmation of GFP and Vio Colonies
  • 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

Saturday, October 15