Team:Caltech/Protocols

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<p>'''Enrichment cultures'''<br/>
<p>'''Enrichment cultures'''<br/>
-
*For BPA (since it was soluble)<br/>
+
* For BPA (since soluble)<br/>
-
1) Set up 16 tubes: 8 tubes vitamin mix vs. 8 tubes no vitamin mix, 4 tubes each of the four locations.<br/>
+
1) Set up 16 tubes: 8 tubes with vitamin media vs. 8 tubes with media (no vitamin), 4 tubes each of the four locations.<br/>
-
2) Place 8 test tubes in 30°C shaker and 8 test tubes in room temperature shaker.</p>
+
2) Place 8 test tubes in 30°C shaker and 8 test tubes in room temperature shaker.<br/>
 +
* For 17α-estradiol, bisphenol A, and nonylphenol (since non-soluble)<br/>
 +
1) Set up two flasks: one with vitamin media, one without vitamin.<br/>
 +
2) Add small amounts (around 50mL or 50mg) of the ten LA river samples into each flask.</p>
}}
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Revision as of 09:03, 22 June 2011


Caltech iGEM 2011



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Recipes for Mixes . Basic Preparation

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 soluble)
1) Set up 16 tubes: 8 tubes with vitamin media vs. 8 tubes with media (no vitamin), 4 tubes each of the four locations.
2) Place 8 test tubes in 30°C shaker and 8 test tubes in room temperature shaker.
  • For 17α-estradiol, bisphenol A, and nonylphenol (since non-soluble)
1) Set up two flasks: one with vitamin media, one without vitamin.
2) Add small amounts (around 50mL or 50mg) of the ten LA river samples into each flask.


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