Team:Cornell/Week 20

From 2011.igem.org

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(Wednesday, October 19)
(Wednesday, October 19)
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Afternoon lab work done by: James Mathew, Charlie Chung, Youjin Cho
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Afternoon lab work done by: Jim Mathew, Charlie Chung, Youjin Cho
:'''Lysing GFP and pZE12 control'''
:'''Lysing GFP and pZE12 control'''
::*Lysed the GFP and pZE12 control using BugBuster.
::*Lysed the GFP and pZE12 control using BugBuster.

Revision as of 22:56, 21 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 |


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°C 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°C oven overnight

Tuesday, October 18

Afternoon lab work done by: Bill Jo

Making Microfluidic Chips
  • Cut out PDMS and bonded to glass slide using O2 plasma
  • Tested resulting chips, 3 new chips made
  • Poured 55g of 10:1 PDMS over mold and put in the 60°C oven overnight

Wednesday, October 19

Morning lab work done by: Bill Jo

Making Microfluidic Chips
  • Cut out PDMS and bonded to glass slide using O2 plasma
  • Tested resulting chips, 8 new chips

Afternoon lab work done by: Jim Mathew, Charlie Chung, Youjin Cho

Lysing GFP and pZE12 control
  • Lysed the GFP and pZE12 control using BugBuster.
  • After adding 5ml of BugBuster to each samples, shook the samples for 20 minutes.
  • Spun the samples down for 20 minutes at 4°C at maximum speed.
(Lysed samples of GFP and pZE12 control stored in 4°C)

Thursday, October 20

Friday, October 21

Afternoon lab work done by: Archana Rachakonda, Jim Mathew, Charlie Chung

  • Spin down 1L cultures of newly transformed (because of sequencing results negative for PCR deletion) VioA, B, E and pZE12
- Stored in -20°C freezer of Olin 301
  • PCR of GFP-AviTag-pZE12 to create final construct of iGEM prefix-GFP-AviTag-iGEM suffix
  • Run on gel, extract, and purify
  • Digest with EcoRI and PstI

Saturday, October 22