title: A microplate assay for K-ras genotyping.
abstract: The paper presents a microplate hybridisation assay for the detection of codon 12 mutations of the K-ras protooncogene. Single-stranded target DNA, obtained from amplifying sample DNA with 5'-biotin and 5'-digoxigenin-labelled primers and subsequent strand separation, is hybridised with solid phase-fixed capture probes complementary to wild-type and mutated forms of K-ras. After stringent washing the duplex DNA is detected by an ELISA-like protocol incorporating photometric, fluorometric or luminometric detection. Application examples are shown in which the K-ras status was examined in peripheral blood cells, cell cultures, fresh and paraffin-embedded tumour tissue and in stool samples.
authors.affiliation: Institut für Pathologische und Klinische Biochemie, Berlin-Buch, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Berndt
authors.forename: G
authors.initials: G
authors.lastname: Wolf
authors.forename: G
authors.initials: G
authors.lastname: Schröder
authors.forename: M
authors.initials: M
authors.lastname: Bebenroth
authors.forename: K
authors.initials: K
authors.lastname: Oehlschlegel
authors.forename: T
authors.initials: T
authors.lastname: Hillebrand
authors.forename: J
authors.initials: J
authors.lastname: Zanow
mesh_terms: D003062:Codon / Q000737:chemistry; D004252:DNA Mutational Analysis / Q000379:methods; D011905:Genes, ras / Q000235:genetics*; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
chemical_list: D003062:Codon