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. 1992 Jul;13(1):116-23.

A two-step hybridization method for chemiluminescent detection of single copy genes

Affiliations
  • PMID: 1503762

A two-step hybridization method for chemiluminescent detection of single copy genes

Q Nguyen et al. Biotechniques. 1992 Jul.

Abstract

We have developed a technique for the chemiluminescent detection of single copy genes that eliminates the high backgrounds and problems with probe labeling associated with existing methods. The procedure employs a primary hybridization of single-stranded probe DNA to immobilized target DNA, a secondary hybridization with a covalently cross-linked oligonucleotide-alkaline phosphatase conjugate, followed by incubation in the chemiluminescent substrate AMPPD and detection on x-ray film. The key to the success of this method is that the primary probe contains a region complementary to the target DNA as well as to the oligonucleotide sequence of the secondary probe-alkaline phosphatase conjugate. Here we report our results using the two-step hybridization procedure to detect single copy genes from genomic Southern blots.

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