Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/09: Difference between revisions
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(Autocreate 2012/11/09 Entry for Haynes_Lab:Notebook/Investigating_Photo-Switchable_Synthetic_Nucleosomes) |
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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 ∞ | |||
<center> | |||
[[Image:VN_Gel_11-9-12.png|150px|Results of the digests are shown.]] | |||
[[Image:KAH_Fermentas_GeneRuler_1kbplus.jpg]] | |||
</center> | |||
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Revision as of 05:15, 12 November 2012
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November 9, 2012
PCR program:
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