RNase protection/PCR product

Kathleen Hopp khopp at POST.ITS.MCW.EDU
Mon Dec 13 19:12:49 EST 1993


Dear Bionetters,
     I am attempting to implementt RNase protection to verify expression
of a PCR product obtained by RT-PCR of material from cell culture. My
protection probe (even after gel purification) always has multipke (maybe
multitudinous is a better word) smaller bands. I've tried a couple changes
in procedure but am ready to go all the why back to making new DEPC water.
    Anyone have any advice or experience to offer? Altermatively is there
another way to check if my PCR product is truly a reflection of a species
present in the culture material? We have sequenced it and it reflects a 
"double (read that  double) exon product where a alternative splicing
situation is usually found.

    Please excuse the noise... it is not expletives!

   Thank you in advance,



        Kathie Hopp   Medical College of Wisconsin



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