High-specificity in-situ hybridization. Methods and application.
We describe a technique of in-situ hybridization using oligonucleotide probes employing the expression of immunoglobulin VH genes as a model. Optimal conditions for hybridization with the 35S-labeled oligonucleotide probes were established with monoclonal B-cell lines that express VH genes of known nucleic acid sequence. The range of sensitivity and specificity achieved with this technique is documented. Under conditions of high stringency, this method can detect the expression of highly related VH hypervariable regions.
Long
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AA
Department of Medicine, Tufts University School of Medicine, New England Medical Center, Boston, MA 02111.
Mueller
J
J
Andre-Schwartz
J
J
Barrett
K J
KJ
Schwartz
R
R
Wolfe
H
H
D001402:B-Lymphocytes / Q000276:immunology; D001483:Base Sequence; D002999:Clone Cells / Q000276:immunology; D004247:DNA / Q000235:genetics; D005069:Evaluation Studies as Topic; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain; D005803:Genes, Immunoglobulin; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods* / Q000706:statistics & numerical data; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000235:genetics; D012680:Sensitivity and Specificity
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D004247:DNA