Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/12: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(2 intermediate revisions by one other user not shown) | |||
Line 7: | Line 7: | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
==November 12, 2012== | ==November 12, 2012== | ||
* The PCR products were purified with the Zymo DNA Clean | * The PCR products were purified with the Zymo DNA Clean & Concentrator Kit | ||
{| class="wikitable" border=1 cellpadding="5" cellspacing="0" | {| class="wikitable" border=1 cellpadding="5" cellspacing="0" | ||
Line 17: | Line 17: | ||
| LOV || 0.048 || 0.026 || 1.860 || 48.095 | | 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 [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 || --- || --- || --- | |||
|- | |||
| 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 | |||
Revision as of 05:51, 15 November 2012
Project name | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||
November 12, 2012
Great! Next step...
|