bacterioplankton communities in the Danjiangkou Reservoir

A
total of 35 surface-water samples (seven replicates for each sample) were
collected from the Danjiangkou Reservoir on May 1, 2016. Large plankton were
removed by filtering through a 20-μm membrane filter, and bacterioplankton were
collected by filtering through a 0.22-μm-diameter filter. The genomic DNA of
bacterioplankton was extracted from the stored filters using the E.Z.N.A.®
Water DNA Kit (OMEGA, USA) according to the manufacturer's instructions.
Electrophoresis and a NanoDrop ND 2000 (Thermo Scientific, USA) were used to
examine the quantity of the extracted DNA. The V3-V4 region of the bacterial
16S rRNA gene was amplified using 338F and 806R with sample-identifying
barcodes.High-throughput sequencing (Illumina MiSeq PE300 platform, Illumina,
USA) was performed by Shanghai Majorbio Bio-Pharm Technology Co., Ltd. (Shanghai,
China).

2016年5月1日，从丹江口水库共采集35份地表水样本，每份样本设置7个平行重复样。通过20μm孔径滤膜过滤去除大型浮游生物，再经0.22μm孔径滤膜收集浮游细菌。采用美国OMEGA公司的E.Z.N.A.® Water DNA试剂盒，依照制造商提供的操作说明书，从保存的滤膜上提取浮游细菌的基因组DNA。通过电泳法与NanoDrop ND 2000超微量核酸蛋白测定仪（Thermo Scientific，美国）对提取得到的DNA进行定量检测。使用带有样本识别条码的引物338F和806R，扩增细菌16S rRNA基因的V3-V4高变区。本次高通量测序由中国上海的上海美吉生物医药科技有限公司，基于Illumina MiSeq PE300测序平台（Illumina，美国）完成。