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    Nucleic Acids Res. 2009 Oct;37(19):6291-304. Epub 2009 Sep 16.

    Multiplex primer prediction software for divergent targets.

    Source

    Computations/Global Security, Lawrence Livermore National Laboratory and QuantaLife, Inc, Livermore, CA, USA. gardner26@llnl.gov

    Abstract

    We describe a Multiplex Primer Prediction (MPP) algorithm to build multiplex compatible primer sets to amplify all members of large, diverse and unalignable sets of target sequences. The MPP algorithm is scalable to larger target sets than other available software, and it does not require a multiple sequence alignment. We applied it to questions in viral detection, and demonstrated that there are no universally conserved priming sequences among viruses and that it could require an unfeasibly large number of primers ( approximately 3700 18-mers or approximately 2000 10-mers) to generate amplicons from all sequenced viruses. We then designed primer sets separately for each viral family, and for several diverse species such as foot-and-mouth disease virus (FMDV), hemagglutinin (HA) and neuraminidase (NA) segments of influenza A virus, Norwalk virus, and HIV-1. We empirically demonstrated the application of the software with a multiplex set of 16 short (10 nt) primers designed to amplify the Poxviridae family to produce a specific amplicon from vaccinia virus.

    PMID:
    19759213
    [PubMed - indexed for MEDLINE]
    PMCID:
    PMC2770652
    Free PMC Article

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