Microbiota of intestinal contents of dissected mice. mouse gut metagenome

Whole intestines of 6 mice were dissected into 6 sections (SI1, SI2, SI3, SI4, Cecum, LI). The intestinal tract contents were washed out using 1ml of PBS buffer and subjected to enzyme-based DNA extraction process. The 16S rRNA gene hypervariable V1-V2 regions were PCR-amplified using universal primers (a barcoded 27Fmod and 338R) and, multiplexed amplicon pyrosequencing was carried out using 454 GS FLX Titanium or 454 GS JUNIOR (Roche Applied Science).

将6只小鼠的完整肠道解剖分割为6个区段（SI1、SI2、SI3、SI4、盲肠（Cecum）、大肠（LI））。使用1ml磷酸盐缓冲液（PBS buffer）冲洗肠道内容物，随后开展基于酶解法的DNA提取流程。采用通用引物（带条形码标记的27Fmod与338R）对16S rRNA基因的高变V1-V2区域进行PCR扩增，并借助454 GS FLX Titanium或454 GS JUNIOR测序平台（罗氏应用科学（Roche Applied Science））完成多重扩增子焦磷酸测序。