Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms.

Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms.
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DOI:
10.1038/ismej.2012.8
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发表时间:
2012-08
期刊:
The ISME journal
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--
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其他
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随着Illumina HiSeq2000在10天的运行中产生高达600 GB的成对端100碱基读取,DNA测序的成本继续下降。在这里,我们提出了一种在HiSeq2000和MiSeq Illumina平台上进行群落扩增子测序的方法,并将该方法应用于来自寄主相关环境和自由生活环境的24个微生物群落的测序。随着更多测序平台的出现,一个关键的问题是,在一个平台上得出的生物学结论是否与在另一个平台上得出的结论一致。我们表明,为这些仪器开发的方法成功地重新获得了已知的生物学结果,此外,生物学结论在测序平台(HiSeq2000与MiSeq)和扩增片段的测序区域之间是一致的。
DNA sequencing continues to decrease in cost with the Illumina HiSeq2000 generating up to 600 Gb of paired-end 100 base reads in a ten-day run. Here we present a protocol for community amplicon sequencing on the HiSeq2000 and MiSeq Illumina platforms, and apply that protocol to sequence 24 microbial communities from host-associated and free-living environments. A critical question as more sequencing platforms become available is whether biological conclusions derived on one platform are consistent with what would be derived on a different platform. We show that the protocol developed for these instruments successfully recaptures known biological results, and additionally that biological conclusions are consistent across sequencing platforms (the HiSeq2000 versus the MiSeq) and across the sequenced regions of amplicons.
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