User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/04/11: Difference between revisions
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> Digests (Fermentas FD)<br> | |||
# | BD-112 and BD-113 | ||
[[Image:BD112-BD113-Gel.JPG|thumb|350px|2 colonies of BD-112 (1 & 2) and 2 colonies of BD-113 (3 & 4)- White dashed lines border, 1 and 2 show the vector backbone (3426 bp) and the BD-112 constructed gene(T7 + RBS + PcTF + 2 scars) (1196 bp), 3 and 4 show the vector backbone (2079 bp) and the BD-113 constructed gene(T7 + RBS + PcTF + 2 scars) (1196 bp).]] | |||
{| class="wikitable" width=400px | |||
| Plasmid DNA || 2.0 μl* | |||
|- | |||
| EcoR1 || 1.0 μl | |||
|- | |||
| Pst1 || 1.0 μl | |||
|- | |||
| 10x FastDigest buffer + green loading dye || 1.5 μl | |||
|- | |||
| dH<sub>2</sub>O || 9.5 μl | |||
|- | |||
| || 15.0 μl total | |||
|- | |||
| Incubate at 37°C for 10 minutes. | |||
|} | |||
# Make the (1%) agarose glee and add 15μl of restricted DNA (PcTF & RBS) in one well and 10 μl of ladder. | |||
* The size of the constructed gene supposed to be: 1123 + 15 + 6 + 6 + 46 = 1144 bp' |
Revision as of 13:57, 11 April 2012
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11/4/12> Digests (Fermentas FD)
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