Please help me to discuss Southern Hyb. result

angie kookkui at care2.com
Thu Feb 28 01:42:11 EST 2002


I use 650 bp probe for Southern Hyb. with genomic DNA (complete digestion)
in low stringency temp.
The result that I've got is my 2 genomic samples have nice band with
background. But 1 genomic sample has "dot" band....it doesn't like band but
just a dot in size equally as the 2 genomic samples. Is that "dot" count to
be a band?
and another question. My positive control in this Hyb, got 2 bands even I
expected just 1. The size around 3 kb. but another unexpected band is 10 kb.
Is that the uncomplete digestion? 'cause I load much more genomic DNA.
My positive control is E. coli JM109. Is it possible that the carbohydrate
will obstruct the EcoR1 so it happened the uncomplete digestion so the hyb.
band occured?
Please help me to discuss that.
Thank you. 



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