User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/03/05: Difference between revisions
From OpenWetWare
(Autocreate 2013/03/05 Entry for User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli) |
(→Date) |
||
| Line 7: | Line 7: | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
= | =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 15:35, 5 March 2013
 PcTF Subcloning in E. coli
|
<html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||
3/5/2013PcTF + His Tag Ligation in V0120 PCR
|
|||||||||||||||||
