PMID: 8962112Dec 10, 1996Paper

Peptide nucleic acid pre-gel hybridization: an alternative to southern hybridization

Proceedings of the National Academy of Sciences of the United States of America
H Perry-O'KeefeM Egholm

Abstract

We have found that it is possible to use labeled peptide nucleic acid (PNA)-oligomers as probes in pre-gel hybridization experiments, as an alternative for Southern hybridization. In this technique, the PNA probe is hybridized to a denatured DNA sample at low ionic strength and the mixture is loaded directly on to an electrophoresis system for size separation. Ensuing gel electrophoresis separates the single-stranded DNA fragments by length. The neutral backbone of PNA allows for hybridization at low ionic strength and imparts very low mobility to excess PNA. Detection of the bound PNA is possible by direct fluorescence detection with capillary electrophoresis, or the DNA/PNA hybrids can be blotted onto a membrane and detected with standard chemiluminescent techniques. Efficient single bp discrimination was achieved routinely using both capillary and slab-gel electrophoresis.

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Citations

May 10, 2006·Applied Microbiology and Biotechnology·Shantanu Karkare, Deepak Bhatnagar
Nov 1, 1988·Molecular and Cellular Endocrinology·Y TakuwaH Rasmussen
Apr 17, 1990·Molecular and Cellular Endocrinology·W S Zawalich, H Rasmussen
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