Rumen metagenome of 437 cows. METALGEN

We sequence the metagenome of 437 Holstein cows using nanopore sequencing. Genomic DNA was extracted from 250 µl of each thawed and homogenized ruminal content sample, using the “DNeasy Power Soil” commercial kit (QIAGEN, Valencia, CA, USA). 260/280 and 260/230 ratios were around 1.8 and 2.0, respectively. Oxford Nanopore Technologies (ONT) SQK-LSK109 Ligation Sequencing kit was used for multiplexed sequencing in MinION automatic sequencer. 1D Native barcoding genomic DNA (EXP-NBD104 or EXP-NBD114) ONT kit was used for sample multiplexing, pooling barcoded DNA from 12 samples for each run. Pooling was done using a 1.5 ml DNA LoBind tube to perform adapter ligation using a R9.4.1 flow cell.

本研究采用纳米孔测序技术，对437头荷斯坦奶牛的宏基因组进行了测序。从每份解冻并均质化的瘤胃内容物样本中取250微升，使用商用"DNeasy Power Soil"试剂盒（凯杰公司，QIAGEN，美国加利福尼亚州巴伦西亚）提取基因组DNA，其260 nm/280 nm与260 nm/230 nm吸光度比值分别约为1.8和2.0。本次实验使用牛津纳米孔科技公司（Oxford Nanopore Technologies, ONT）的SQK-LSK109连接测序试剂盒，在MinION全自动测序仪上开展多重测序。采用ONT的1D原生条形码基因组DNA试剂盒（EXP-NBD104或EXP-NBD114）进行样本多重标记，每次测序运行均将12个带有条形码的DNA样本进行混合。样本混合操作在1.5毫升DNA低吸附管中完成，随后使用R9.4.1测序流动槽完成接头连接反应。