Virginia Commonwealth/4 August 2009

From 2009.igem.org

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Contents

Tuesday 4 August 2009

Results

Maria and Afton

  • Overnight culture of I1352 plated on AMP:
    • Sample taken from plate after PSB4C5 ligation attempt successfully grew expressing RFP. The color of the sample was pink
    • Sample taken from plate after ligation with itself did not grow at all, and expressed no RFP
  • There is a possibility the pSB4C5 backbone may be a bad part
    • previous years it has not functioned properly
    • Gel electrophoresis indicates DNA Gel Purification may be necessary
  • Possible inefficiency of BioBrick enzymes may have come from repeated freezing and thawing
    • They will be aliquoted into smaller amounts
  • Gel Electrophoresis Results
From Left to Right: 100bp Ladder, Bioline Hyperladder, I1352(Digest), I1352(Miniprep), pSB4C5(Digest), pSB4C5(Miniprep)

Trentay 22:25, 4 August 2009 (UTC)


Tasks

Maria and Afton

  • Run a Gel Electrophoresis
  • Transform and grow up pSB1C3 with the death gene P1010

Trentay 22:11, 4 August 2009 (UTC)


Wetlab

  • New BioBrick Assembly Kit came in and the following were aliquoted into their respective amounts
Name Amount/Tube # Tubes Extra in Stock
XbaI 10 µL 15 No
SpeI 5 µL 10 No
PstI 10 µL 25 Yes
EcoRI-HF 10 µL 25 Yes
BSA 5 µL 8 Yes
T4 Ligase Buffer 10 µL 25 Yes
T4 Ligase 10 µL 8 No
  • To show how many times a tube has been thawed and refrozen, a hash mark will be placed on the side of the tube, each time it is thawed as a reference for others to see

Trentay 15:24, 5 August 2009 (UTC)

Maria and Afton

  • Gel Electrophoresis was run
    • pSB4C5 and I1352 digested and undigested (right after Miniprep) were run on a gel and results showed excessive bands in the undigested DNA. DNA Gel Purification will be done on future Miniprepped DNA
  • pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer.
  • Transformation was done using DB 3.1
Name Plate Growth Observation
pSB1C3 CM n/a
(+) Control LB n/a
(+) Control zapped LB n/a
(-) Control CM n/a

Trentay 22:33, 4 August 2009 (UTC)