Mutational analysis of human NRAS genes in malignant melanoma: Rapid methods for oligonucleotide hybridization and manual and automated direct sequencing of products generated by the polymerase chain reaction Journal Article

  

Authors:	Dicker, A. P.; Volkenandt, M.; Albino, A. P.
Article Title:	Mutational analysis of human NRAS genes in malignant melanoma: Rapid methods for oligonucleotide hybridization and manual and automated direct sequencing of products generated by the polymerase chain reaction
Abstract:	Three methods to detect single base mutations in codon 61 of the human NRAS gene from human melanoma DNA are described and compared: oligonuceotide hybridization analysis and direct manual and automated sequence analysis. Point mutations are detected by oligonucleotide hybridization and direct manual and direct automated sequence analysis of in vitro amplified genomic DNA. Heterozygosity for mutant alleles is reliably detected by oligonucleotide hybridization and by direct manual, but not by direct automated, sequence analysis. Generating single‐stranded DNA via “asymmetric polymerase chain reaction (PCR)” and utilizing α35S‐dATP as radiolabel for manual sequencing and fluorescent‐dye labeled primers for automated sequencing (Applied Biosystems, Inc.), we can obtain sequence information from either strand. The use of several of these methodologies to detect single base changes in the human NRAS gene is illustrated. In addition, the use of these and other related techniques to define the involvement of RAS oncogenes in human melanomas more precisely is reviewed. Copyright © 1990 Wiley‐Liss, Inc., A Wiley Company
Keywords:	gene mutation; review; dna polymerase; polymerase chain reaction; melanoma; fluorescent dye; tumor cells, cultured; automation; time factors; molecular sequence data; isotope labeling; dna, neoplasm; base sequence; dna mutational analysis; genes, ras; nucleic acid hybridization; dna, single-stranded; oligonucleotide; oligodeoxyribonucleotides; hybridization; human; support, non-u.s. gov't; support, u.s. gov't, p.h.s.
Journal Title:	Genes Chromosomes and Cancer
Volume:	1
Issue:	4
ISSN:	1045-2257
Publisher:	Wiley Periodicals, Inc
Date Published:	1990-03-01
Start Page:	257
End Page:	269
Language:	English
DOI:	10.1002/gcc.2870010402
PUBMED:	2278957
PROVIDER:	scopus
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-0025370201&doi=10.1002%2fgcc.2870010402&partnerID=40&md5=5fa13dd84b446d77554b615b93a11106
Notes:	Source: Scopus

https://synapse.mskcc.org/synapse/works/has_related_data_chk/159308