The oligomer extension" hot blot": a rapid alternative to Southern blots for analyzing polymerase chain reaction products.

JD Parker, GC Burmer - Biotechniques, 1991 - europepmc.org
JD Parker, GC Burmer
Biotechniques, 1991europepmc.org
We report a modification of a liquid hybridization method that serves as a rapid, sensitive
alternative to Southern hybridization for the analysis of polymerase chain reaction (PCR)
products. An aliquot of the completed PCR is mixed with an internally nested, end-labeled
oligonucleotide probe, and one cycle of PCR is performed. The products are
electrophoresed, dried and analyzed by autoradiography. The method is significantly more
sensitive than standard Southern hybridization methods, is equally specific, requires as little …
We report a modification of a liquid hybridization method that serves as a rapid, sensitive alternative to Southern hybridization for the analysis of polymerase chain reaction (PCR) products. An aliquot of the completed PCR is mixed with an internally nested, end-labeled oligonucleotide probe, and one cycle of PCR is performed. The products are electrophoresed, dried and analyzed by autoradiography. The method is significantly more sensitive than standard Southern hybridization methods, is equally specific, requires as little as 1-10 picograms of probe and results can be obtained in less than an hour. The procedure is equally useful for identifying products in multiplex PCRs or single-stranded PCRs.
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