Haynes Lab:Notebook/Inteins/2013/03/01

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