User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/03/05
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| - | = | + | =3/5/2013= |
| - | ''' | + | '''PcTF + His Tag Ligation in V0120''' |
| - | # | + | |
| + | PCR | ||
| + | * 95°C, 3 min. | ||
| + | * [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35 | ||
| + | * 72°C, 3 min. | ||
| + | * 4°C, ∞ | ||
| + | |||
| + | {| {{table}} border="1" cellspacing="3" <!-- PCR rxn. table --> | ||
| + | |- valign="top" | ||
| + | | bgcolor=#cfcfcf | Reagent | ||
| + | | bgcolor=#cfcfcf | Volume | ||
| + | | rowspan="7" | <u>Expected:</u><br>1. H2B = 378<br>2. LOV = 450<br>3, 4. pSB1A3 = ~2000 | ||
| + | | rowspan="7" | [[Image:KAH022313_gel1.jpg|200px|Hover name]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
| + | |- | ||
| + | | DNA(plasmid) || 0.2 μL | ||
| + | |- | ||
| + | | 10 μM primer 1 || 1.0 | ||
| + | |- | ||
| + | | 10 μM primer 2 || 1.0 | ||
| + | |- | ||
| + | | 2x GoTaq || 25.0 | ||
| + | |- | ||
| + | | dH<sub>2</sub>O || 22.8 | ||
| + | |- | ||
| + | | || 50.0 μL | ||
| + | |} | ||
| + | |||
| + | |||
| + | Purification | ||
| + | * Clean with Zymo DNA c&c; elute w/ 20 μL dH<sub>2</sub>O | ||
| + | * Measure [DNA] | ||
Revision as of 18:35, 5 March 2013
 PcTF Subcloning in E. coli
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3/5/2013PcTF + His Tag Ligation in V0120 PCR
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