User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/01/12: Difference between revisions
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Ligation of KAH111, KAH112 and C-His vector in DH5α-T and BL21 | |||
* The 4 μL of C-His Vector from stock is diluted with 20 μL of nuclease free water, then cut with E/P and purified from the gel. | |||
* Ligations | |||
{| {{table}} cellspacing="3" <!-- Ligations table --> | |||
|- bgcolor=#cfcfcf | |||
| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/12/12</font> | |||
|- | |||
| 1. E. coli DH5α-T KAH111(E/P)/size, 20 ng + C-His(E/P)/3600, 26ng || <font color="blue">KAH111/C-His 3:1 No Colonies</font> | |||
|- | |||
| 2. E. coli BL-21 KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng || <font color="blue">KAH111/C-His 3:1 No Colonies</font> | |||
|- | |||
| 3. C-His(E/P)/ 25 ng || | |||
|} | |||
{| {{table}} cellspacing="3" <!-- Ligation rxn table --> | |||
| || 1 || 2 || 3 | |||
|- | |||
| Insert DNA || 3.12 || 3.12 || --- | |||
|- | |||
| Vector DNA || 2.94 || 2.94 || 2.94 | |||
|- | |||
| 2x lgn buf (Roche) || 7.06 || 7.06 || 5.0 | |||
|- | |||
| T4 ligase (NEB) || 1.0 || 1.0 || 1.0 | |||
|- | |||
| dH<sub>2</sub>O || 0 || 0 || 0.88 | |||
|- | |||
| || 14.12 μL || 14.12 μL || 10μl | |||
|} |
Revision as of 13:57, 12 January 2013
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01/12/2013Ligation of KAH111, KAH112 and C-His vector in DH5α-T and BL21
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