User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/03/05: Difference between revisions
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''' | '''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. KAH109 = 1132 <br>2. KAH111 = 1132 <br>3. KAH112 = 1132 <br>4. KAH115 = 931 <br>5. KAH148 = 967 <br> 6. KAH150 = 967 <br> 7. KAH151 = 967 | |||
| rowspan="7" | [[Image:PCR KAH Product.png|400px|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 16:19, 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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