complete ligation HELP?! - (Jul/04/2006 )
I have read a couple conversations on different forms of plasmid...and I understand that the nicked circle and relaxed circle are both products of ligation. My question is ...are there any ways to make the ligation complete....into a covalently closed (supercoiled) plasmid....AND
can the nicked and relaxed plasmids transform into bacteria successfully???
My following step transfection into mammalian cells also requires a closed plasmid I believe...or at least that is the best form for it.
Ames
-Ames-
QUOTE (Ames @ Jul 4 2006, 04:32 PM)
I have read a couple conversations on different forms of plasmid...and I understand that the nicked circle and relaxed circle are both products of ligation. My question is ...are there any ways to make the ligation complete....into a covalently closed (supercoiled) plasmid....AND
can the nicked and relaxed plasmids transform into bacteria successfully???
My following step transfection into mammalian cells also requires a closed plasmid I believe...or at least that is the best form for it.
Ames
can the nicked and relaxed plasmids transform into bacteria successfully???
My following step transfection into mammalian cells also requires a closed plasmid I believe...or at least that is the best form for it.
Ames
Okay I am replying to myself...because I realize now that ligation results in closed relaxed and nicked circles that become repaired in the proper host.
-Ames-