Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/09
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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 08:15, 12 November 2012
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November 9, 2012
PCR program:
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