Team:UNITN-Trento/Notebook/Labposts/06/13

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{ "date" : "2013-06-12", "author" : "Gabriele", "title" : "Ligation and transformation", "content" : "Ligation of SAMsynthetase and pSB1C3 with and without RFP, so I prepared four samples:
1) pSB1C3 with RFP (Ctrl_1)
2) pSB1C3 with RFP + SAMsynthetase (1:2)
3) pSB1C3 without RFP (Ctrl_2)
4) pSB1C3 without RFP (1:2)
The samples were prepared and ligation was performed following the ligation protocol.

Ligation parameters
  • Plasmid concentration: 27.8ng/µl
  • Plasmid length: 2070bp
  • Insert concentration: 13.2ng/µl
  • Insert length: 1155bp
  • Used plasmid: 250µl
  • Volume of reaction: 35µl
  • Buffer concentration: 10X

Transformation in NEB10β cells was performed following the transformation protocol.Plates in incubator ON at 37°C static, more than 16 hours.", "tags" : "SAMsynthetase" }