Metagenome from Paralichthys olivaceus

Food-poisoning fillet of Paralichthys olivaceus. Total DNA or RNA was prepared from a 30 mg of the frozen fillet of Paralichthys olivaceus by RecoverAll Total Nucleic Acid isolation kit (Ambion, Austin, TX USA). Double-stranded cDNA was prepared from 1 micro g of total RNA using the random priming method with SuperScript Choice System for cDNA synthesis (Invitrogen, Carlsbad, CA, USA). cDNA was purified using a QIAquick PCR Purification kit (QIAGEN, Hilden, Germany). Short-read DNA sequencing using the Illumina Genome Analyzer II An approximately 100-bp length DNA library was prepared from the mixture of 2 micro g of total DNA and 0.5 micro g of cDNA using a genomic DNA sample prep kit (Illumina, San Diego, CA, USA), and DNA clusters were generated on a slide using a Single Read Cluster Generation kit (ver. 2) on an Illumina cluster station (Illumina), according to the manufacturer's instructions.

致食物中毒的牙鲆（Paralichthys olivaceus）鱼片样本。采用RecoverAll总核酸分离试剂盒（Ambion，美国德克萨斯州奥斯汀市），从30 mg冷冻牙鲆鱼片中提取总DNA（total DNA）或总RNA（total RNA）。使用SuperScript cDNA合成首选系统（Invitrogen，美国加利福尼亚州卡尔斯巴德市），以随机引物法从1 μg总RNA中合成双链cDNA（double-stranded cDNA）。使用QIAquick PCR纯化试剂盒（QIAGEN，德国希尔德市）对cDNA进行纯化。采用Illumina Genome Analyzer II开展短读长DNA测序（short-read DNA sequencing）：依照制造商操作说明，首先使用基因组DNA样本制备试剂盒（Illumina，美国加利福尼亚州圣迭戈市），将2 μg总DNA与0.5 μg cDNA混合后构建约100 bp的DNA文库（DNA library）；随后使用单端读取簇生成试剂盒（版本2）在Illumina簇仪的载玻片上生成DNA簇（DNA clusters）。