< Haynes Lab:Notebook | Inteins | 2013 | 03
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- 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.
- 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