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Descripción
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| Construction of a microorganism culture collections isolated from environment can be time- consuming since individual colonies containing single microbes might demand several streaking steps for colony purification. In addition, the pursuing of pure colonies may restricts important insights on mutual association. Potentially, in extreme cases, microorganisms in consortia might depend strictly on beneficial microbe-microbe interactions. Here we present a method for construction and identification of a collection of microorganisms and consortia of microorganisms isolated by a single step of plating and colony picking. Complex microbial mixtures isolated from sugarcane roots and stalks were plated in defined media at proper dilution to obtain colonies. Colonies were picked, grown in 96-deepwell plates and stored at - 80 °C. This collection comprises over five thousand wells containing individual microorganisms or consortia. In order to identify these microorganisms we have developed a method to sequence pools of amplicons in 96-well plates using barcodes to tag plates, rows and columns in two PCR steps. Pooled plates can be sequenced into a single run of PacBio and delivers high quality amplicons relying on circular consensus sequence (CCS) for error corrections. Together, the multi-tagging system and PacBio platform allows the identification of microbe composition of each well, regardless if it contains pure or consortia colonies. Results will be discussed in the context of cost-effective alternatives for large-scale identification of individuals in complex colony communities by sequencing pools of multiplexed 96-well plates | |
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Internacional
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Si |
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Nombre congreso
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Rhizosphere 4 |
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Tipo de participación
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970 |
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Lugar del congreso
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Maastricht |
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Revisores
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Si |
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ISBN o ISSN
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0000-0000 |
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DOI
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Fecha inicio congreso
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20/06/2015 |
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Fecha fin congreso
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25/06/2015 |
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Desde la página
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380 |
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Hasta la página
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380 |
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Título de las actas
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- |