User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/04/18: Difference between revisions
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'''Building a Reporter Gene to Determine Chromatin Protein Function''' | '''Building a Reporter Gene to Determine Chromatin Protein Function''' | ||
*Ligation of KAH182 = BBa_S04739 and KAH201 = BBa_S04745 and cloning in DH5α-T. | *Ligation of KAH182 = BBa_S04739 and KAH201 = BBa_S04745 and cloning in DH5α-T. | ||
*Assembly strategy: The backbone KAH201(S/P)/3403 , the insert is KAH182(X/P)/1675. | *Assembly strategy: The backbone KAH201(S/P)/3403 , the insert is KAH182(X/P)/1675. | ||
=12/1/12= | |||
> Digests (Fermentas FD)<br> | |||
[[Image:Vector_Cut_(E-P).png |thumb|350px| Human Vector PT7CFE1-CHis, 3600 bp, Cut with EcoRI and PstI Separately.]] | |||
{| class="wikitable" width=400px | |||
| Plasmid DNA || 30.00 μl | |||
|- | |||
| Enzyme 1* || 1.0 μl | |||
|- | |||
| Enzyme 2** || 1.0 μl | |||
|- | |||
| 10x FastDigest buffer + green loading dye || 1.5 μl | |||
|- | |||
| dH<sub>2</sub>O || 0.00 μl | |||
|- | |||
| || 30.0 μl total | |||
|- | |||
| Incubate at 37°C for 10 minutes. | |||
|} | |||
* Enzyme 1 for KAH182 is Xba1 and for KAH201 is Spe1. | |||
** Enzyme 2 for KAH182 and KAH201 is Pst1. | |||
# Make the (1%) agarose glee and add 15μl of restricted vector in one well and 10 μl of ladder. | |||
Revision as of 22:11, 18 April 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> | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
04/18/2013Building a Reporter Gene to Determine Chromatin Protein Function
12/1/12> Digests (Fermentas FD)
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