i got colonies back. I did miniprep to see whether my insert is in it. i will do digestion later. generally do you run the plasmid from miniprep to see the size difference? my insert is 200bp, 300bp. could i be able to tell from the gel of plasmids?
by the way, i do feel i like here. people are really friendly and helpful!!
Thanks friends
cathy
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2 replies to this topic
#2
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Posted 30 March 2005 - 07:33 AM
hi
yep i do minipreps to check plasmids and presence of insert.
But if you have many colonies you can try pcr on colonies
fred
look at amberg lab protocol.
yep i do minipreps to check plasmids and presence of insert.
But if you have many colonies you can try pcr on colonies
fred
look at amberg lab protocol.
#3
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Posted 30 March 2005 - 01:09 PM
I try to do a definitive test digest before I run things on a gel.
Circular and supercoiled plasmids migrate at a different rate in agarose compared to linear DNA. Granted you can still tell apart empty vectors from those containing inserts, but you're running a gel anyway, better get the most info from one gel run, right?
Circular and supercoiled plasmids migrate at a different rate in agarose compared to linear DNA. Granted you can still tell apart empty vectors from those containing inserts, but you're running a gel anyway, better get the most info from one gel run, right?
Edited by george@CASE, 30 March 2005 - 01:09 PM.
