Immersion-histo polymerase chain reaction: a practical tool for visualization of single-copy genes in tissue sections.
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.
Pan
L
L
Department of Histopathology, University College London Medical School, United Kingdom.
Diss
T C
TC
Peng
H
H
Isaacson
P G
PG
D017931:DNA Primers; D019286:Endopeptidase K / Q000378:metabolism; D016591:Histocytological Preparation Techniques; D006801:Humans; D017403:In Situ Hybridization; D008198:Lymph Nodes / Q000473:pathology; D016393:Lymphoma, B-Cell / Q000235:genetics / Q000473:pathology; D008224:Lymphoma, Follicular / Q000235:genetics / Q000473:pathology; D008867:Microtomy; D014066:Palatine Tonsil / Q000473:pathology; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D019286:Endopeptidase K