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Team:Cornell/Week 20
From 2011.igem.org
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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
Night lab work done by: Nicholas Kramer
- Making Microfluidic Chips
- Poured 55g of 10:1 PDMS over the molds and put in 60 degree Celsius oven overnight
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
Afternoon lab work done by: Maneesh Gupta and Claire Paduano
- Making Microfluidic Chips
- Cut PDMS off of mold and bonded to glass slide with O2 plasma
- Tested resulting chips, 6 new chips made
- Poured 55g of 10:1 PDMS on molds and put in the 60 degree Celsius oven overnight
