Team:Caltech/Protocols

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Revision as of 22:47, 21 June 2011

Caltech iGEM 2011

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Transforming DNA:
1) Thaw competent cells on ice: 15K, 15J, 15O, 15M.
2) Get 10 micro Liters of pure water.
3) Pipette out DNA from the source plate and put it in the appropriate tubes.
4) Flick the competent cells in the tube gently.
5) Pipette 40 micro Liters of competent cells into the DNA.
6) Pipette 2 micro Liters of the DNA and put it in the competent cell tubes.
7) Stir a bit.
8) Leave on ice for 30 minutes.
9) Heat Shock for 45 sec by using a water bath set to 42°C and then thawing on ice for 2 min.
10) Pipette 500 micro Liters of S.O.C. (LB + glucose) into each competent cell tubes.

Enrichment cultures

For BPA (since it was soluble)

1) Set up 16 tubes: 8 tubes vitamin mix vs. 8 tubes no vitamin mix, 4 tubes each of the four locations.
2) Place 8 test tubes in 30°C shaker and 8 test tubes in room temperature shaker.

@@ Line 19: / Line 19: @@
 ) Set up 16 tubes: 8 tubes vitamin mix vs. 8 tubes no vitamin mix, 4 tubes each of the four locations.<br/>
 ) Place 8 test tubes in 30°C shaker and 8 test tubes in room temperature shaker.</p>
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