Premium
Investigation of a quantitative post‐hybridization signal amplification system for mRNA‐oligodeoxyribonucleotide in situ hybridization
Author(s) -
Hoyland Judith A.,
Freemont Anthony J.
Publication year - 1991
Publication title -
the journal of pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.964
H-Index - 184
eISSN - 1096-9896
pISSN - 0022-3417
DOI - 10.1002/path.1711640110
Subject(s) - biotinylation , in situ hybridization , streptavidin , oligonucleotide , microbiology and biotechnology , in situ , molecular probe , nucleic acid thermodynamics , biotin , messenger rna , chemistry , biology , biochemistry , dna , base sequence , gene , organic chemistry
This study was undertaken to assess a post‐hybridization amplification system for increasing the sensitivity of in‐situ hybridization with oligodeoxyribonucleotide (oligonucleotide) probes. The technique employs a multilayer streptavidin–biotinylated link protein amplification technique, similar to that used in the ABV histochemical amplification system, to attach increasing numbers of radiolabelled streptavidin molecules to a biotinylated ologonucleotide probe. In tissue sections the amplification technique increases the signal‐to‐noise ratio dramatically, increasing the sensitivity of in situ hybridization and reducing the autoradiographic exposure time. On an artificial medium the amplification technique has been shown to increase the hybridization signal 100‐fold when compared with a directly radiolabelled oligonucleotide probe. The technique could be used to compare relative amounts of target molecule, there being a linear relationship between the detectable signal and the log of the concentration of the target molecule.