Haynes Lab:Notebook/Engineering PC-TFs/2014/03/20: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Line 25: | Line 25: | ||
| || 12.0 μL total || same | | || 12.0 μL total || same | ||
|- | |- | ||
| colspan="3" | Mix the reaction(s) thoroughly by flicking the tube.<br>Incubate at room temperature for 10 minutes. | | colspan="3" | Mix the reaction(s) thoroughly by flicking the tube.<br>Incubate at room temperature for 10 minutes. Repeat once again for BL21. 6μL of buffer was used because the total volume with no water exceeded 10μL. | ||
|} | |} | ||
# Calculate how many ng of insert you need to get a 2:1 ratio of insert molecules to 50 ng vector molecules<br>''X ng CMV = (bp CMV = 588 / bp MV9 = 5291) x 2 x 50 ng MV9 = 11.1132ng insert (CMV)'' | # Calculate how many ng of insert you need to get a 2:1 ratio of insert molecules to 50 ng vector molecules<br>''X ng CMV = (bp CMV = 588 / bp MV9 = 5291) x 2 x 50 ng MV9 = 11.1132ng insert (CMV)'' |
Revision as of 15:52, 20 March 2014
<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> | |||||||||||||||||||||||||
Summary
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.
'Transforming bacteria with the ligated plasmids in BL21'
|