Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/13: Difference between revisions
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(Autocreate 2012/11/13 Entry for Haynes_Lab:Notebook/Investigating_Photo-Switchable_Synthetic_Nucleosomes) |
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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 [http://partsregistry.org/Part:pSB1A2 pSB1A2 plasmid] (from Rene) XbaI and SpeI | |||
{| class="wikitable" border=1 cellpadding="5" cellspacing="0" | |||
|- | |||
| 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 | |||
|- | |||
| dH<sub>2</sub>O || 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 | |||
* Extract DNA from gel | |||
Revision as of 21:19, 13 November 2012
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November 13, 2012
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