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American Journal of Pathology, Vol 150, 1245-1252, Copyright © 1997 by American Society for Investigative Pathology

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Immersion-histo polymerase chain reaction: a practical tool for visualization of single-copy genes in tissue sections

L Pan, TC Diss, H Peng and PG Isaacson
Department of Histopathology, University College London Medical School, United Kingdom.

A method, immersion-histo polymerase chain reaction (IH-PCR), was developed for visualization of single-copy DNA sequences in cells in paraffin-embedded tissue sections. Sections were mounted on coverslips, cut into small pieces, and immersed in reaction mixtures in micro-tubes for specific DNA amplification using a conventional thermal cycler. This was followed by in situ hybridization, in micro-tubes, with PCR- generated, digoxigenin-labeled probes. Epstein-Barr virus, chromosomal translocations, and rearranged immunoglobulin genes were clearly demonstrated in individual cells in human lymphomas. The use of a standard thermal cycler, the ease of probe preparation, and the high specificity and sensitivity of target localization make IH-PCR a practical and powerful tool for the study of genetic events within individual cells.




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Copyright © 1997 by the American Society for Investigative Pathology.