A modified nick translation method used with FISH that produces reliable results with archival tissue sections.
Nick translation is used to label DNA and RNA to produce probes for in situ hybridization and Northern and Southern blotting. Fluorescence in situ hybridization (FISH) is a widely applied technique used to determine chromosomal and genetic anomalies in many biological samples. Initially the technique was applied to metaphase preparations, but the usefulness of detecting genetic anomalies in solid tumors in situ has resulted in the development of modified protocols. Formalin fixed paraffin processed tissue sections present novel challenges when applying FISH; the probes must be small (between 200 and 600 base pairs) and pretreatment is necessary before the probes can be applied to tissue sections, to promote probe access to target DNA. Here we report on a modification of a nick translation method to produce a probe that can reliably be used with FISH in paraffin processed tissue sections.
Watters
Amanda D
AD
University Dept of Surgery, Glasgow Royal Infirmary, Scotland, UK.
Stacey
Michael W
MW
Bartlett
John
J
D015152:Blotting, Northern; D015139:Blotting, Southern; D002898:Chromosomes, Human, Pair 8 / Q000648:ultrastructure; D003360:Cosmids / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism
D012313:RNA; D004247:DNA; D003850:Deoxyribonuclease I