Team:Stony Brook/Notebook/Cancer-W1

Cancer Group: Week 1

Week 1 (6/27 - 7/2)




6/27

Project Progress

  • 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

Interlab Study

  • Measured LUDOX and H2O for plate reader
  • Transformation of positive and negative controls, test devices 1-3



6/28

Project Progress

  • 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

Interlab Study

  • Heat and "SB" streaking seems not to have worked
  • No visible red colonies on plate



6/29

Project Progress

  • PCR Not working well → keep trying new methods with different annealing temperatures
  • Tried 65 and 67 degrees C

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

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



7/1

  • 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



7/2

  • Ran yesterday's digestion on gel