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

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===DTL of Wintergreen pathway===
===DTL of Wintergreen pathway===


* Digested 1 microgram of J45004 and 20ng of J45017
* Digested 400ng of J45004, 20ng of J45017, and 1 microgram of B21 (for v0120 backbone)
** Digested J45004, J45017, and B21 BB with XbaI/PstI
* Gel extracted and purified bands indicated in 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
 




Revision as of 06:44, 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


VP16 PCR 2

No PCR product is visible


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
  • Gel extracted and purified bands indicated in 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



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