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

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November 12, 2012 The PCR products were purified with the Zymo DNA Clean & Concentrator 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