Team:Cornell/Week 20

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Results | Protocol | Notebook | Parts Submitted

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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

Tuesday, October 18

Wednesday, October 19

Thursday, October 20

Friday, October 21

Saturday, October 22