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Chemiluminescent detection of DNA probes in forensic analysis
Author(s) -
Klevan Leonard,
Horton Liz,
Carlson David P.,
Eisenberg Arthur J.
Publication year - 1995
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501601258
Subject(s) - chemiluminescence , oligonucleotide , dioxetane , hybridization probe , dna , oligomer restriction , alkaline phosphatase , chemistry , microbiology and biotechnology , chromatography , biology , enzyme , biochemistry
The conditions for hybridization and detection of enzyme‐labeled probes have been optimized in our laboratory for use with oligonucleotides coupled to alkaline phosphatase. We have examined several enzyme‐linked probes which are complimentary to commonly used variable number of tandem repeats (VNTR) loci to determine the feasibility of using chemiluminescence for routine application in forensic DNA analysis. It was found that a chemiluminescent detection system employing an alkaline phosphatase activated dioxetane in the presence of chemiluminescent enhancers provides a high degree of sensitivity in hybridization protocols with a significant savings in overall filter processing time. The chemiluminescent system achieved equal or greater sensitivity than observed for 32 P‐labeled probes in much shorter development times. Furthermore, a new chemiluminescent substrate, Lumi‐Phos® Plus, has recently been investigated and found to further decrease the filter development time for forensic assays.