Team:Caltech/Protocols

From 2011.igem.org

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<p>'''Transforming DNA:'''<br/>  
<p>'''Transforming DNA:'''<br/>  
1) Thaw competent cells on ice: 15K, 15J, 15O, 15M.<br/>  
1) Thaw competent cells on ice: 15K, 15J, 15O, 15M.<br/>  
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8) Leave on ice for 30 minutes.<br/>  
8) Leave on ice for 30 minutes.<br/>  
9) Heat Shock for 45 sec by using a water bath set to 42°C and then thawing on ice for 2 min.<br/>
9) Heat Shock for 45 sec by using a water bath set to 42°C and then thawing on ice for 2 min.<br/>
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10)Pipette 500 micro Liters of S.O.C. (LB + glucose) into each competent cell tubes.<br/>   
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10)Pipette 500 micro Liters of S.O.C. (LB + glucose) into each competent cell tubes.<br/>  </p>
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(STILL CONFUSED ABOUT THIS PROTOCOL NOT SURE IF RIGHT)</p>
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<p>'''I don't know what this is called'''...Putting 100 mg soil into each tube, then put the tubes in shakers overnight at 30°C and room temperature respectively???</p>
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<p>'''I don't know what this is called'''
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...Putting 100 mg soil into each tube, then put the tubes in shakers overnight at 30°C and room temperature respectively???</p>
<p>'''Starter Competent Cell Culture Recipe?'''<br/>  
<p>'''Starter Competent Cell Culture Recipe?'''<br/>  
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[[Team:Caltech/Recipes|Recipes for Mixes]]
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Revision as of 22:36, 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.

I don't know what this is called ...Putting 100 mg soil into each tube, then put the tubes in shakers overnight at 30°C and room temperature respectively???

Starter Competent Cell Culture Recipe?
1) Take the competent cell tubes out of the freezer.
2) Vortex it and take 100 micro Liters from it and pipette it into the labeled plates.
3) Put in some beads and shake gently.
4) Put the dirty beads away in the dirty bead container.


Recipes for Mixes

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