southern blot - gel loading - (May/02/2005 )

how much digested genomic dna from mammalian cells should i load into a gel to see single copy gene in southern blot? i am using dig probe and alreadyi tried 5 micrograms, however i haven't saw anything...
thank you

have you checked your probe has been labelled with DIG? We ahve found that it is incorporated quite lousy.

A good way to check is to spot your probe on a corner of a membrane and see if that lights up.

Good luck

Nick

Current protocols says that 10 micrograms is the max that a nylon membrane will hold. hope that helps.

-mike-