PCR probes for Southern blot - (Apr/24/2006 )
Hi
I want to detect series of stress related genes in various species of same genus . I am planning to do Southren blot for that. Is it OK if I develop a probe using PCR with raidolabelled dNTPS , Is any one has done it?. Because most of them use radom primer method to develop probes , is there any reason for that, if it is so please let me know.If any one has simple lab stardized protocl for Southern blot , please do send me
Thanking you all
shashi
-shashi-
QUOTE (shashi @ Apr 24 2006, 01:29 PM)
Hi
I want to detect series of stress related genes in various species of same genus . I am planning to do Southren blot for that. Is it OK if I develop a probe using PCR with raidolabelled dNTPS , Is any one has done it?. Because most of them use radom primer method to develop probes , is there any reason for that, if it is so please let me know.If any one has simple lab stardized protocl for Southern blot , please do send me
Thanking you all
shashi
I want to detect series of stress related genes in various species of same genus . I am planning to do Southren blot for that. Is it OK if I develop a probe using PCR with raidolabelled dNTPS , Is any one has done it?. Because most of them use radom primer method to develop probes , is there any reason for that, if it is so please let me know.If any one has simple lab stardized protocl for Southern blot , please do send me
Thanking you all
shashi
Probes can be generated by PCR, we use this procedure pretty frequently in our lab (althoug we use DIG label not radiolabeling [High Prime Kit from Roche]). Two reasons a random primer method may be used is that either you don't have sequence data to design primers or the fragment is too long/difficult to amplify by PCR.
-arthrobacter-