LAFLAMME@MAINE.BITNET (10/26/90)
Does anyone out there in netland have any experience in the use of oligonucleotide probes to screen total DNA in northern or slot-blots? I have been using 20 mers to pick out individual members of a family of plant genes. It seems that the method is not sensitive enough to detect these sequences in a sample containing a HUGE excess of non-hybridizing species. If anyone could pass along a helpful hint I would be very happy. I would prefer not to isolate poly A RNA due to the extra hassle and the possibility of skewing the final results.
mkw6d@krebs.acc.Virginia.EDU (Martyn K. White) (10/30/90)
In article <90298.171656LAFLAMME@MAINE.BITNET> LAFLAMME@MAINE.BITNET writes: >Does anyone out there in netland have any experience in the use of >oligonucleotide probes to screen total DNA in northern or slot-blots? >I have been using 20 mers to pick out individual members of a family >of plant genes. It seems that the method is not sensitive enough to (stuff deleted) Have you thought of PCR. Martyn K. White, Box 441, U.Va, Cville VA 22908 (804) 924 8710 mkw6d@krebs.acc.Virginia.EDU mkw6d@Virginia.EDU
IBT329%DJUKFA11@PUCC.PRINCETON.EDU (11/05/90)
It might be that asymmetric PCR is an answer: i.e. Primer A = 50 - 100 pmol Primer B = 0.5 - 2 pmol Bilanovic D.D