User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/04/22: Difference between revisions
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''' | '''Building a Reporter Gene to Determine Chromatin Protein Function (Ligation into the mammalian vector)''' | ||
*Ligation of construct KAH182-KAH201 and mammalian vector V0200 and cloning in DH5α-T.. | |||
KAH182 = BBa_S04739 and KAH201 = BBa_S04745 and V0200 = BBa_J176121. | |||
*Assembly strategy: The backbone KAH201(S/P)/3403 , the insert is KAH182(X/P)/1675. | |||
'''Digests''' <br> | |||
[[Image:KAH201-KAH182 Ligation.jpg|thumb|400px| (1,2) KAH182, 1675 bp, Cut with Xba1 and Pst1, (3,4) KAH201, 3403 bp, Cut with Spe1 and Pst1]] | |||
{| {{table}} cellspacing="3" width=350px | |||
| Plasmid DNA || 25.00 μl | |||
|- | |||
| NotI || 1.0 μl | |||
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| SpeI || 1.0 μl | |||
|- | |||
| 10x FastDigest buffer + green loading dye || 3.00 μl | |||
|- | |||
| dH<sub>2</sub>O || 0.00 μl | |||
|- | |||
| || 30.0 μl total | |||
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| Incubate at 37°C for 10 minutes. | |||
|} | |||
Revision as of 15:38, 22 April 2013
 PcTF Subcloning in E. coli
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DateBuilding a Reporter Gene to Determine Chromatin Protein Function (Ligation into the mammalian vector)
KAH182 = BBa_S04739 and KAH201 = BBa_S04745 and V0200 = BBa_J176121.
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