Haynes Lab:Notebook/Engineering PC-TFs/2012/11/26

From OpenWetWare
Jump to: navigation, search

Hayneslab3.gif
Asu logo 3.gif

Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Summary

  • Digest the pSB1A2/3 plasmid (from Rene) with XbaI & SpeI to get rid of the insert
Reagent Volume  
DNA (plasmid) 20.0
10x buffer 3.0
XbaI 1.0
SpeI 1.0
dH2O 5.0
  30 μL --> 37°C/ ~10 min.
  • Run entire 30 μL on a 1% gel (use big tooth comb) next to a ladder
  • You should see two bands. The larger band (2000 bp) is the backbone.
  • Cut out and gel purify this fragment (ignore the shorter fragment).
  • Notify Dr. Haynes when you are done. The next step is Gibson Assembly and transformation.