技術文獻
技術文獻
Faecal microbiota transplantation‐mediated jejunal microbiota changes halt high‐fat diet‐induced obesity in mice via retarding intestinal fat absorptionIF:1.40
The jejunal mucosal scrapings were collected and immediately snap-frozen in liquid nitrogen and stored at 80°C.The QIAamp DNA stool Mini Kit (Qiagen, Hilden, Germany) was used to extract DNA from enteric bacteria injejunal mucosal scrapings and PCR amplification of theV3-V4 region (338F: ACTCCTACGGGAGGCAGCA,806R: GGACTACHVGGGTWTCTAAT) of 16S rDNA. Theamplification products were detected by 2% gel electrophoresis and PCR products were recovered by axyprepDNA gel extraction kit (Axygen, San Francisco, CA, USA).The PCR products were detected and quantified byQuantiFluorTM-ST microfluorometer (Promega, Madison,WI, USA) and then Miseq library was constructed forpyrosequencing on the Illumina MiSeq platform (Illumina,Madison, WI, USA). Clean sequences were assigned to
the same operational taxonomic units (OTUs) based on asimilarity ≥ 97%. Data were analysed on the free onlinecloud platform (https://cloud.majorbio.com/) of Majorbio(Shanghai, China). Bacterial species were further identi-fied by sequence comparison of OTUs on NCBI. The relative abundance of significantdifferences in phylum, genusand OTU levels was examined by the Kruskal–Wallis ranksum test. Spearman’s correlation analysis was furtheranalysed to uncover the relationship between jejunalmicrobiota and intestinal fat absorption.
感謝動物營養與飼料科學重點實驗室(華東),農業農村部,浙江杭州 310058,中國。浙江省動物飼料與營養重點實驗室,浙江杭州 310058,中國。浙江大學預防獸醫研究所,浙江杭州 310058,中國。6浙江省預防獸醫重點實驗室,引用文獻!
- 上一條:如何避免外部溫度對細胞或活體組織的影響
- 下一條:如何判斷PCR檢測試劑盒的批間差異
