Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/13

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(Autocreate 2012/11/13 Entry for Haynes_Lab:Notebook/Investigating_Photo-Switchable_Synthetic_Nucleosomes)
Current revision (07:48, 15 November 2012) (view source)
(November 13, 2012)
 
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==Entry title==
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==November 13, 2012==
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* Insert content here...
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* Digests:
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** In a 30 μL reaction, cut H2B with XbaI and SpeI
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** In a 30 μL reaction, cut LOV with 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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{| class="wikitable" border=1 cellpadding="5" cellspacing="0"
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|-
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| Reagent || H2B|| LOV || pSB1A2
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|-
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| DNA || 8.0 || 8.0 || 15.0
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|-
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| XbaI || 1.0|| 1.0 || 1.0
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|-
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| SpeI || 1.0 || 1.0 || 1.0
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|-
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| 10x buffer || 3.0 || 3.0 || 3.0
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|-
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| dH<sub>2</sub>O || 10.0 || 10.0 || 10.0
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|-
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| Total vol. || 23. 0 || 23.0  || 30.0
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|}
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* Run entire 30 μL on a 1% gel (use the big fat-tooth well comb)
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* Cut band out of gel (use UV box); H2B = ~410, LOV = ~461, pSB1A2 = ~2000
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<center>
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[[Image:VN_Gel_11-14-12.png|200px|Results of from the digests are shown.]]
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[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]]
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</center>
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<center>
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Results of the enzyme digests are shown, with H2B on the left, LOV in the middle, and pSB1A3 on the right. The gel was loaded with 30 μL samples.  The bands that were cut are outlined in green.
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</center>
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November 13, 2012

  • 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 10.0 10.0 10.0
Total vol. 23. 0 23.0 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

Results of from the digests are shown. Image:KAH_Fermentas_GeneRuler_1kbplus.jpg

Results of the enzyme digests are shown, with H2B on the left, LOV in the middle, and pSB1A3 on the right. The gel was loaded with 30 μL samples. The bands that were cut are outlined in green.



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