Department of Computer Science, Northern Arizona University, Flagstaff, AZ, USA.
ISME J. 2012 Aug;6(8):1621-4. doi: 10.1038/ismej.2012.8. Epub 2012 Mar 8.
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.
DNA 测序的成本持续降低，Illumina HiSeq2000 每运行 10 天可生成多达 600GB 的配对末端 100 碱基读长。在此，我们展示了在 HiSeq2000 和 MiSeq 高通量测序平台上进行群落扩增子测序的方案，并将该方案应用于从宿主相关和自由生活环境中提取的 24 个微生物群落。随着更多测序平台的出现，一个关键问题是在一个平台上得出的生物学结论是否与在不同平台上得出的结论一致。我们表明，为这些仪器开发的方案成功地再现了已知的生物学结果，并且在测序平台（HiSeq2000 与 MiSeq）和扩增子测序区域上，生物学结论是一致的。
