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

AMERICAN JOURNAL OF PATHOLOGY(1997)

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Abstract
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, digoxigenein-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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Key words
immunoglobulins,dna,in situ hybridization,virus,sequence,translocation,localization,genes,polymerase chain reaction,dna sequence,genetics,cell,gene,microtomy,dna primers
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