Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/12: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(Autocreate 2012/11/12 Entry for Haynes_Lab:Notebook/Investigating_Photo-Switchable_Synthetic_Nucleosomes) |
(fix raw html notebook nav) |
||
(6 intermediate revisions by 2 users not shown) | |||
Line 2: | Line 2: | ||
|- | |- | ||
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### 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== | ||
* | * The PCR products were purified with the Zymo DNA Clean & Concentrator Kit | ||
{| class="wikitable" border=1 cellpadding="5" cellspacing="0" | |||
|- | |||
| 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 [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 | |||
Latest revision as of 22:14, 26 September 2017
Project name | Main project page Previous entry Next entry | |||||||||||||||||||||||||||||||||||||||||||
November 12, 2012
Great! Next step...
|