A microplate assay for K-ras genotyping
Berndt, C.; Wolf, G.; Schröder, G.; Bebenroth, M.; Oehlschlegel, K.; Hillebrand, T.; Zanow, J.
European Journal of Clinical Chemistry and Clinical Biochemistry 34(10): 837-840
1996
ISSN/ISBN: 0939-4974 PMID: 8933108 Document Number: 456783
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 turnout tissue and in stool samples.