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Descripción
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| Unbiased identification of organisms by PCR reactions using universal primers followed by DNA sequencing assumes positive amplification. We used six universal loci spanning 48 plant species and quantified the bias at each step of the identification process from end point PCR to next-generation sequencing. End point amplification was significantly different for single loci and between species. Quantitative PCR revealed that Cq threshold for various loci, even within a single DNA extraction, showed 2,000- fold differences in DNA quantity after amplification. Nextgeneration sequencing (NGS) experiments in nine species showed significant biases towards species and specific loci using adaptor-specific primers. NGS sequencing bias may be predicted to some extent by the Cq values of qPCR amplification | |
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Internacional
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JCR del ISI
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Título de la revista
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Analytical And Bioanalytical Chemistry |
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ISSN
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1618-2642 |
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Factor de impacto JCR
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3,578 |
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Información de impacto
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Volumen
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407 |
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DOI
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Número de revista
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Desde la página
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1841 |
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Hasta la página
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1848 |
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Mes
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SIN MES |
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Ranking
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