Team:Stony Brook/Notebook/Week1

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Week 1: 6/27-7/1

Week 1 (6/27 - 7/2)

6/27

Cancer Group
  • PCR of 3xHA-TDGF1 construct. 12 cycles
  • Gel of PCR Product → PCR was a failure. Band was 300bp. Should have been 750bp
  • Second PCR of 3xHA-TDGF1 construct. 25 cycles instead of 12 → left overnight
Vaccine Group
  • Things
Interlab Study
  • Measured LUDOX and H2O for plate reader
  • Transformation of positive and negative controls, test devices 1-3

6/28

Cancer Group
  • Ran gel on PCR of yesterday's construct → Gel worked
  • Gel purification of PCR Product → low concentration of DNA found during nanodrop
  • PCR the construct using a revised method
  • 2:54pm → ran the gel on the PCR product
Vaccine Group
  • Things
Interlab Study
  • Heat and "SB" streaking seems not to have worked
  • No visible red colonies on plate

6/29

Cancer Group
  • PCR Not working well → keep trying new methods with different annealing temperatures
  • Tried 65 and 67 degrees C
Vaccine Group
  • Things
Interlab Study
  • FITC started
  • Cutting re-inoculated
  • Ordered new LUDOX
Other
  • College of Arts and Sciences Pre College Summer Institute students stopped by lab
  • Spoke to them about information on synthetic biology and iGEM

6/30

Cancer Group
  • PCR worked → Test 5 → changed annealing temperature to 66 degrees C + increased the denaturing time
  • Nanodropped DNA
Vaccine Group
  • Things

7/1

Cancer Group
  • Digestion of TDGF-1 construct and YFP352GAP vector with two restriction enzymes
  • Will ligate later
TDGF-1 Construct (Phire PCR 128.2 ng/mL) YEP352GAP Vector (343.2 ng/mL)
Total Reaction Volume 50 uL 50 uL
Nuclease Free Water 27.4 uL 37.2 uL
10x Cutsmart Buffer 5 uL 5uL
xho1 1 uL 1 uL
Pvu II 1 uL 1 uL
DNA 15.6 uL 5.8 uL
Vaccine Group
  • Things

7/2

Cancer Group
  • Running yesterday's digestion on gel
Vaccine Group
  • Stuff & Things