A simple DNA diagnostic method for human genetic disorders |
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Authors: | Daniel Rabin Nanibhushan Dattagupta |
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Affiliation: | (1) Molecular Diagnostics, Inc., 400 Morgan Lane, 06516 West Haven, CT, USA |
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Abstract: | Summary A fast, reliable, and simple technique for detecting point mutations in unfractionated human DNA has been developed. Oligonucleotide probes complementary to either sickle- or normal -globin DNA are labeled by primer extension and hybridized to DNA applied to nitrocellulose paper in a dot-blot format. A short hybridization time (about 1 h) and low probe concentration (about 1 nM) yield low background and high specificity. Double-blind trials show 100% agreement with restriction fragment length polymorphism (RFLP) analysis of DNA from normal, sickle, and heterozygous subjects. |
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