Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/12

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** In a 30 μL reaction, cut H2B with XbaI and SpeI
** In a 30 μL reaction, cut H2B with XbaI and SpeI
** In a 30 μL reaction, cut LOV with XbaI and SpeI
** In a 30 μL reaction, cut LOV with XbaI and SpeI
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** In a 30 μL reaction, cut a pSB1A2 plasmid (from Rene) XbaI and SpeI
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** In a 30 μL reaction, cut a [http://partsregistry.org/Part:pSB1A2 pSB1A2 plasmid] (from Rene) XbaI and SpeI
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Revision as of 19:33, 13 November 2012

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November 12, 2012

  • The PCR products were purified with the Zymo DNA Clean and Concentration Kit
PCR Product 260 280 260/280 ng/μL
H2B 0.052 0.028 1.865 51.607
LOV 0.048 0.026 1.860 48.095


Great! Next step...

  • Digests:
    • In a 30 μL reaction, cut H2B with XbaI and SpeI
    • In a 30 μL reaction, cut LOV with XbaI and SpeI
    • In a 30 μL reaction, cut a pSB1A2 plasmid (from Rene) XbaI and SpeI
Reagent H2B LOV pSB1A2
DNA 8.0 8.0 15.0
XbaI 1.0 1.0 1.0
SpeI 1.0 1.0 1.0
10x buffer 3.0 3.0 3.0
dH2O --- --- ---
Total vol. 30. 0 30.0 30.0


  • Run entire 30 μL on a 1% gel (use the big fat-tooth well comb)
  • Cut band out of gel (use UV box); H2B = ~410, LOV = ~461, pSB1A2 = ~2000
  • Extract DNA from gel



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