User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/10/10: Difference between revisions
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* '''Restriction enzyme deactivation''' Incubate the tubes at 80°C for 10 minutes and cool down gradually in room temperature. | * '''Restriction enzyme deactivation''' Incubate the tubes at 80°C for 10 minutes and cool down gradually in room temperature. | ||
'''Ligate The Sticky Ends to Make Circular DNA''' | *'''Ligate The Sticky Ends to Make Circular DNA''' | ||
{| {{table}} cellspacing="3" width=350px | |||
| DNA Conc. || 10ng || 20ng || 30ng || 50ng || 50ng (Ctrl) | |||
|- | |||
| DNA μL || 1μL || 2μL || 3μL || 5μL || 5μL | |||
|- | |||
| T4 Ligase || 1.0 μl || 1.0μL || || 1.0μL || 1.0μL || 0.0μL | |||
|- | |||
| 2X Roche Buffer|| 5.00 μl || 5.00 μL || 5.00 μL || 5.00 μL || 5.00 μL | |||
|- | |||
| dH<sub>2</sub>O || 5.00 μl || 5.00 μl || 5.00 μL || 6.00 μL || 0.00 μL | |||
|- | |||
| Total || 10.0 μl || 10.0 μl || 10.0 μl || 10.0 μl || 10.0 μl | |||
|- | |||
| Incubate at room temperature for 30 minutes. | |||
|} |
Revision as of 18:10, 13 October 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> | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
10/10/12Site Directed Mutagenesis-Removing the BSMBI cut site from BD002 reporter gene
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