Difference between revisions of "IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/08/02"

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(DTL of Mira/Brazz+StrepII+Stop into V24)
(Gel Purification)
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*Gel bands indicated were cut and purified from the gel.
  
 
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Revision as of 10:13, 3 August 2010

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Team Fence

Gel extraction

Gal4 PCR product and Ec digest gel extract and purified.

Gal on the left, EcR on the right


Gal4 Ecr digest gel ex 8-2.jpg

VP16 PCR 2

No PCR product is visible

VP16PCR.2 blank 8-2.jpg


Team Flavor

DTL of Mira/Brazz+StrepII+Stop into V24

  • Digested 1 microgram of DNA for each insert and for V24 backbone.
    • Digested inserts and backbone with NotI/SpeI
  • Gel extracted and purified bands indicated in image below
  • Ligated insert into 50ng of V24 backbone with a 3:1 insert to backbone ratio
  • Transformed and plated on LB+AMP plates and left in 37°C incubator overnight

DTL of Wintergreen pathway

  • Digested 400ng of J45004, 20ng of J45017, and 1 microgram of B21 (for v0120 backbone)
    • Digested J45004, J45017, and B21 BB with XbaI/PstI
  • PCR purified digested inserts
  • Gel extracted and purified B21 BB (see gel image below)
  • Ligated inserts into 50ng of B21 v0120 backbone with a 3:1 insert to backbone ratio
    • Transformed and plated on LB+AMP plates and left in 37°C incubator overnight

Gel Purification

Mbv24b21digest.jpg

  1. Ladder
  2. Mira N SS
  3. Mira C SS
  4. Brazz N SS
  5. Brazz C SS
  6. V24 N/S
  7. B21 X/P
  8. Ladder
  • Gel bands indicated were cut and purified from the gel.