Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/09: Difference between revisions
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|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}}}}]]}} | ||
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== | ==November 9, 2012== | ||
* | |||
* First, resuspend dry primer oligos (in blue-capped tubes) to 100 μM | |||
* Then make working solutions of 10 μM | |||
* Reactions: | |||
# H2B-GFP plasmid + BB_H2B fwd + BB_H2B rev | |||
# LOV plasmid + BB_LOV fwd + BB_LOV rev | |||
{| class="wikitable" border=1 cellpadding="5" cellspacing="0" | |||
|- | |||
| Reagents || H2B || LOV | |||
|- | |||
| Plasmid DNA || 0.5 μL || 0.5 | |||
|- | |||
| primer 1 (10 μM) || 1.0 || 1.0 | |||
|- | |||
| primer 2 (10 μM) || 1.0 || 1.0 | |||
|- | |||
| 2x GoTaq mix || 12.5 || 12.5 | |||
|- | |||
| dH<sub>2</sub>O || 10.5 || 10.5 | |||
|} | |||
PCR program: | |||
* 95°C, 3 min. | |||
* [95°C, 30 sec; 57°C, 30 sec.; 72°C, 30 sec.] x35 cycles | |||
* 72°C, 3 min. | |||
* 4°C ∞ | |||
{| class="wikitable" border=0 | |||
|- | |||
| <u>Expected:</u><br>1. H2B = 410 <br>2. LOV = 461 | |||
| <br> | |||
|} | |||
<center> | |||
[[Image:Gel_11-9-12.png|Results of the digests are shown.]] | |||
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]] | |||
</center> | |||
<center> | |||
Results of the PCR product digests are shown, with H2B on the left and LOV on the right. The gel was loaded with 5 μL samples. | |||
</center> | |||
Latest revision as of 22:14, 26 September 2017
Project name | Main project page Previous entry Next entry | ||||||||||||||||||||
November 9, 2012
PCR program:
Results of the PCR product digests are shown, with H2B on the left and LOV on the right. The gel was loaded with 5 μL samples.
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