User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/09/06: Difference between revisions
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'''Confirm The BSMB1 Removal''' | |||
[[Image:Ligation Confirmation.jpg|thumb|350px| Two seperate bands in each column show the vector backbone and the insert, 4442 bps and the BD001 (KAH201+KAH182) (1901) constructed gene.Backbone, 1901 bps, Cut with SpeI and NotI. First column: Ladder, columns 1-5 are 3:1 ratio colonies, column6 is 4:1 ratio, column 7 is 5:1 ratio and column 8 is 10:1]] | |||
# | {| {{table}} cellspacing="3" width=400px | ||
|Plasmid Type|| SD-BD002 <sup>1</sup> || SD-BD002|| SD-BD002 || BD002 | |||
|- | |||
| Plasmid DNA || 3.0 μl || 3.0 μl|| 3.0 μl|| 3.0 μl | |||
|- | |||
| BSMB1 || 1.0 μl || 0.0 μL || 0.0 μL || 1.0 μL | |||
|- | |||
| Spe1 || 0.0 μl || 1.0 μL || 0.0 μL || 1.0μL | |||
|- | |||
| 10x FastDigest buffer + green loading dye || 1.5 μl || 1.5 μl || 1.5 μl || 1.5 μl | |||
|- | |||
| dH<sub>2</sub>O || 9.5 μl || 9.5 μL || 10.5 μL || 9.5 μL | |||
|- | |||
| || 15.0 μl total || 15μL || 15μL || 15μL | |||
|- | |||
| Incubate at 37°C for 10 minutes. | |||
|} | |||
1- SD-BD002 means BD002 plasmid after the Site-Directed Mutagenesis. | |||
# Make the (1%) agarose glee and add 15μl of restricted vector in one well and 10 μl of ladder. |
Revision as of 12:49, 9 September 2013
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Confirm The BSMB1 Removal ![]()
1- SD-BD002 means BD002 plasmid after the Site-Directed Mutagenesis.
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