Drosophila genomic southerns

Richard M Kliman rkliman at runet.edu
Wed Apr 9 12:22:08 EST 1997


In article <97097.000239VHWCC at CUNYVM.CUNY.EDU>,
Harshinee Wijesinghe  <VHWCC at CUNYVM.CUNY.EDU> wrote:
>Could somebody tell me an effective, non-radioactive labelling method
>that can be used for the probe in Drosophila genomic southerns? Thanks
>in advance for any help.

We did genomic Southerns in a molecular biology course this year.  We 
used an actin probe derived from Dictyostelium to detect homologoues in 
D. melanogaster.  Dicty actin-8 PCR was labelled with DIG-UTP by random 
priming using the Genius kit.  Using the color reactions (not the 
chemilumiscent reaction), we were able to clearly detect 6 bands in a D. 
melanogaster HindIII digest.  I was surprised that this method worked 
with a genome the size of Drosophila, but it did.

Rich Kliman
Dept. of Biology
Radford University



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