@article{Germer01011999,
author = {Germer, Søren and Higuchi, Russell}, 
title = {Single-Tube Genotyping without Oligonucleotide Probes},
volume = {9}, 
number = {1}, 
pages = {72-78}, 
year = {1999}, 
doi = {10.1101/gr.9.1.72}, 
abstract ={We report the development of a self-contained (homogeneous), single-tube assay for the genotyping of single-nucleotide polymorphisms (SNPs), which does not rely on fluorescent oligonucleotide probes. The method, which we call T
m-shift genotyping, combines allele-specific PCR with the discrimination between amplification products by their melting temperatures (T
m). Two distinct forward primers, each of which contains a 3′-terminal base that corresponds to one of the two SNP allelic variants, are combined with a common reverse primer in a single-tube reaction. A GC-tail is attached to one of the forward allele-specific primers to increase theT
m of the amplification product from the corresponding allele. PCR amplification, T
manalysis, and allele determination of genomic template DNA are carried out on a fluorescence-detecting thermocycler with a dye that fluoresces when bound to dsDNA. We demonstrate the accuracy and reliability ofT
m-shift genotyping on 100 samples typed for two SNPs, and recommend it both as a simple and inexpensive diagnostic tool for genotyping medically relevant SNPs and as a high-throughput SNP genotyping method for gene mapping.}, 
URL = {http://genome.cshlp.org/content/9/1/72.abstract}, 
eprint = {http://genome.cshlp.org/content/9/1/72.full.pdf+html}, 
journal = {Genome Research} 
}