User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/05/22: Difference between revisions
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'''PcTF | '''PcTF Extraction from ''E-coli''''' | ||
'''Transform bacteria with the ligated plasmids''' ''30 minutes'' | '''Transform bacteria with the ligated plasmids''' ''30 minutes'' | ||
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'''BugBuster Protein Extraction''' | '''BugBuster Protein Extraction''' | ||
#Harvest cells from liquid culture by centrifugation at 16000 ×g for 10 minutes. | #Harvest cells from liquid culture by centrifugation at 16000 ×g for 10 minutes. | ||
# | # Weigh the cell pellet. Weigh the same size empty well, zero the balance, remove the empty well and then weigh the well with cell pellet. | ||
# Re-suspend the cell pellet in room temperature BugBuster Reagent by pipetting or gentle vortexing. Using 5ml of reagent per gram of wet cell paste. |
Revision as of 16:48, 24 May 2012
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5/22/2012PcTF Extraction from E-coli Transform bacteria with the ligated plasmids 30 minutes
Note: The negative control will show you the number of “background” colonies so that you can determine whether your transformation worked, or is just the result of vector self-ligation or selection failure. Grow liquid cultures
BugBuster Protein Extraction
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