Haynes Lab:Notebook/Inteins/2013/03/01: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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==Entry title==
==Entry title==
* Insert content here...
* Completed a restriction/digest from really good mini prep!
*Used brand new enzymes for the restriction/ digest
*Made a brand new gel
1KBT  KAH9(186)  KAH9(160)  KAH10 (116)  KAH10 (106) 
*Put 10 of ladder, 30 in other wells.
http://openwetware.org/wiki/Image:3-1-2013_gel.jpg
*Run dna recovery kit
**Melted @ 55.6 degrees C for 6 min., vortex in between
**8uL + 7 water to elute dna in last step
 
*Completed ligation/transformation
*Used both 2x and 10x buffers
*Get competent cells and keep on ice
*Used both buffers to see if lab shared stock Roche ligation buffer was contaminated.
*Used brand new ligase and ligation buffer
*Left ligations for at least 30 mins.
*Add 50 uL of competent cells
 




Latest revision as of 22:31, 26 September 2017

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Entry title

  • Completed a restriction/digest from really good mini prep!
  • Used brand new enzymes for the restriction/ digest
  • Made a brand new gel

1KBT KAH9(186) KAH9(160) KAH10 (116) KAH10 (106)

  • Put 10 of ladder, 30 in other wells.

http://openwetware.org/wiki/Image:3-1-2013_gel.jpg

  • Run dna recovery kit
    • Melted @ 55.6 degrees C for 6 min., vortex in between
    • 8uL + 7 water to elute dna in last step
  • Completed ligation/transformation
  • Used both 2x and 10x buffers
  • Get competent cells and keep on ice
  • Used both buffers to see if lab shared stock Roche ligation buffer was contaminated.
  • Used brand new ligase and ligation buffer
  • Left ligations for at least 30 mins.
  • Add 50 uL of competent cells



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