one end blunt end and other end sticky end-- ligation - vector with one end sticky and other blunt end (Aug/29/2005 )
If your insert and vector have the been cut with the same RE then you should not need to blunt end your overhangs. The insert should clone better if it has one stick end rather than two blunt ends. Also this will ensure that the insert goes in the right way.
Daniel
Use less BigDye mix
-Daniel Tillett-
QUOTE (Daniel Tillett @ Sep 1 2005, 12:24 PM)
If your insert and vector have the been cut with the same RE then you should not need to blunt end your overhangs. The insert should clone better if it has one stick end rather than two blunt ends. Also this will ensure that the insert goes in the right way.
Daniel
Use less BigDye mix
Daniel
Use less BigDye mix
but then will the ligation reaction conditions be the same? i thought the conditions for sticky end cloning will not work well with blunt ends ??
regards
laxmi
-phytoviridae-
Just use the blunt end conditions 
Daniel
DNA sequencing primers
-Daniel Tillett-
QUOTE (Daniel Tillett @ Sep 1 2005, 12:32 PM)
THANK you very much for all the time and suggestions
yours gratefully
laxmi
-phytoviridae-