Pyrosequencing protocol using a universal biotinylated primer for mutation detection and SNP genotyping

Primer Design and Marker Clustering for Multiplex SNP-IT Primer Extension Genotyping Assay using Statistical Modeling

MOTIVATION The optimization of the primer design is critical for the development of high-throughput SNP genotyping methods. Recently developed statistical models of the SNP-IT primer extension genotyping reaction allow further improvement of primer quality for the assay. RESULTS Here we describe how the statistical models can be used to improve primer design for the assay. We also show how to...

Universal SNP genotyping assay with fluorescence polarization detection.

The degree of fluorescence polarization (FP) of a fluorescent molecule is a reflection of its molecular weight (Mr). FP is therefore a useful detection methodfor homogeneous assays in which the starting reagents and products differ significantly in Mr. We have previously shown that FP is a good detection method for the single-base extension and the 5'-nuclease assays. In this report, we describ...

Multiplex universal genotyping using a modified ARMS-PCR protocol.

DNA mutations underlie the pathology of inherited disease. The spectrum of genetic changes resulting in disease is diverse, often encompassing both point (single-nucleotide) and length (large deletions) mutations: cystic fibrosis (2) and the muscular dystrophies (10) are classic examples. Similarly, mutational events implicated in carcinogenesis include both single nucleotide substitutions and ...

Development of a cost-effective pyrosequencing approach for SNP genotyping in barley

SNP genotyping with fluorescence polarization detection.

When a fluorescent molecule is excited by plane polarized light, the fluorescence emitted is also polarized. The degree of fluorescence polarization (FP) detected, under constant temperature and solvent viscosity, is proportional to the molecular weight of the dye molecule. By monitoring the FP of a fluorescent dye, one can detect significant changes in the molecular weight of the molecule with...