Southern Hybridization - about the probe and DNA concentration (Jun/14/2005 )

Hi,

It will be great if somebody here can enlighten me. Usually what is the concentration for the probe and DNA to hybridize??? (How many ng) I will not bother the minimum concentration of them. I just want to know whether my probe can successfully hybridize to the target DNA.

Thanks.

hi!!


For southern blot you only need 5ug of DNA, this is enought, if it was a northen then you need more, around 20 and 30ug of RNA. About the amount of probe you have to use 50ng.

Good luck!!

Upps!! Remember that when you are doing a southern blot you need a diefernt % of formamide than in northern blot, 50 and 20.


Baobabs

I use 10ug DNA across the board.

For probe: I random label (Amersham RediPrime II works well) 25ng PCR product or purified plasmid fragment per protocol. Size range 200-700bp preferred.

Probe blot with 2-3E5cpm/ml

If i want to do reverse dot blot, any one have any idea bout the DNA and probe concentration??