Problems with Southerns

roger morton ttguy at bigpond.com
Tue Jun 1 07:38:10 EST 1999


"gmani at acs.itd.uts.edu.au" wrote in message ...
>To whom it may concern,
>I have been having a lot of trouble in making my southerns work.I am using
>genomic DNA from a number of CHO cell clones with a probe specific for
>insulin labelled with 32P.I ran the probe alongside the other DNA so as to
>have probe vs labelled probe as a means of detecting the problem.The
>autoradiograph revealed a strong signal for the probe itself however no
>other clone came up,not even the positive control.

Maybe you have far too much DNA in the "unlabled probe contol". pg
quantities is the amount you probably should have for a genomic southern.
You can calculate how much you need from the amount of genomic DNA you are
adding to the blot, the size of the Hamster genome and the size of the
"probe" fragment. The choice of restriction enzyme could be important. Try a
few different ones. Also the time of acid treatment of the gel could be
critical.







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