DIG labelling and detection

dna dna at vt.edu
Mon Oct 7 08:04:59 EST 1996


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




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