User:Wilfred J. Poppinga/Notebook/Wilfreds Project/2009/08/25

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Combining MBP-ArsR with inducible promotors

  • Cut plasmids containing promotor with SpeI and PstI
    • 1 μL 10x fast digest buffer
    • 0.5 μL SpeI
    • 0.5 μL PstI
    • 8 μL Vector
  • Incubate 1 h @ 37 °C
  • Purify cut plasmid using PCR clean up kit
  • Cut plasmids containing MBP-ArsR with XbaI and PstI
    • 1 μL 10x fast digest buffer
    • 0.5 μL XbaI
    • 0.5 μL PstI
    • 8 μL Vector
  • Incubate 1 h @ 37 °C
  • Put restriction on 1% Agarose (TBE) gel
    • Cut band of ~1500 bp out
    • Purify using gel purification kit
      • End volume 30 μL
  • Ligation
    • μL ligation buffer
    • 1 μL ligase
    • μL vector
    • μL insert
    • Incubate

to do

  • Watercampus
    • AquaExplorere
    • Wetsus
    • Rest
  • Safety!!
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