Multiplex Pyrosequencing.

Nader Pourmand; Elahe Elahi; Ronald W Davis; Mostafa Ronaghi

doi:10.1093/nar/30.7.e31

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Multiplex Pyrosequencing.

Nader Pourmand·Elahe Elahi·Ronald W Davis·Mostafa Ronaghi

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Abstract

We describe here the development of a new and simple single-tube multiplex Pyrosequencing assay. Genomic DNA or cDNA was employed to PCR amplify region(s) using biotinylated and normal primer(s). Subsequent to capture of PCR products on streptavidin-coated beads, single-stranded DNA separation and hybridization of multiple sequencing primers, Pyrosequencing was performed. The obtained pyrogram resulted in a unique pattern in which the intensity of the signal determined the number of incorporated nucleotide(s). Here, we demonstrate the use of this multiplex Pyrosequencing for single nucleotide polymorphisms genotyping and microbial typing.

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Animals Base Sequence DNA DNA, Viral Drosophila melanogaster Genotype Hepacivirus Humans Molecular Sequence Data Polymorphism, Single Nucleotide Reproducibility of Results Sequence Alignment Sequence Analysis, DNA Sequence Homology, Nucleic Acid Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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Nader Pourmand, Elahe Elahi, Ronald W Davis, Mostafa Ronaghi. 2002-04-01. Multiplex Pyrosequencing.. https://doi.org/10.1093/nar%2F30.7.e31

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