IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/08/02

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
• 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

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