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Team:Cornell/Week 20
From 2011.igem.org
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Morning lab work done by: Jim Mathew, Charlie Chung | Morning lab work done by: Jim Mathew, Charlie Chung | ||
:*Submitted VioA, B, E and GFP for sequencing to confirm PCR deletion | :*Submitted VioA, B, E and GFP for sequencing to confirm PCR deletion | ||
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:'''Preparing VioA, B, E and pZE12 for Protein Extraction''' | :'''Preparing VioA, B, E and pZE12 for Protein Extraction''' | ||
Revision as of 18:09, 17 October 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 |
October 16th - October 22nd
Sunday, October 16
Morning lab work done by: Charlie Chung
- Preparing 100mL Cultures of VioA, B, E and pZE12 Vector
- Inoculate two baffled flasks of (50mL LB + 50µL ampicillin) each with 1mL of yesterday's culture for a 1:50 dilution
- Shake at 37°C for 2.5 hours and check OD reading (if 0.05-0.08, then ready for IPTG-induction)
- OD after 2.5 hours = 0.1xx (VioA), 0.08 (VioB), 0.1xx (VioE) -- too dense again...
- - Next time, check OD after 2 hours when doing a 1:50 subculture
- Induced with 5µL 1M IPTG for 0.1µM addition to a 50mL culture
- Shaking at room temperature for 20+ hours, after which we will pellet the cells for lysis and protein extraction
Monday, October 17
Morning lab work done by: Jim Mathew, Charlie Chung
- Submitted VioA, B, E and GFP for sequencing to confirm PCR deletion
- Preparing VioA, B, E and pZE12 for Protein Extraction
- Spin down the 100mL of each culture into pellets (3700rpm for 30 min)
- - Stored in -20°C fridge of Olin 301
