In article <32571241.340C at netvigator.com>, lumdicks at netvigator.com says...
>>Hi all,
>> I'm now using a 280bp DIG labelled probe in Southern hybridization to
>detect a single copy gene in mammalian genomic DNA. However, the results
>were very unsatifactory even 50ug od DNA were loaded into each lane. No
>positive bands and results obtained so far. I've followed exactly what
>the DIG user manual said, but no help.
>>Any suggestion..??
>>And do any one know if dextran sulfate or PEG can be added and use in
>DIG Southern hybridization??
>>Besides, the efficiency of DNA transfer in Southern was also very low,
>large amount of DNA left in the gel after the process. I suspect the
>problem was due to the use of an excessively heavy weight at the top. Do
>anyone knows if it was really the cause..???
>>Thanks!!
>>Regards,
>Dicken Lum
Are you sure your labeling reaction worked? Have you tested your probe?
-
John R. McQuiston
Molecular Genetics Lab
CMMID VMRCVM
VPI & SU
1410 Prices Fork Rd
Blacksburg VA 24061
