A microplate assay for K-ras genotyping.
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.
Berndt
C
C
Institut für Pathologische und Klinische Biochemie, Berlin-Buch, Germany.
Wolf
G
G
Schröder
G
G
Bebenroth
M
M
Oehlschlegel
K
K
Hillebrand
T
T
Zanow
J
J
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
D003062:Codon