GSM5034309: Hu85; feces metagenome; OTHER
Fecal DNA extract from 5 g feces of human using the QIAamp DNA Stool Mini Kit (Qiagen) according the instruction of manufactory. QIAseq 16S/ITS 96-Index I (384) (Cat# 333825) and QIAseq 16S/ITS Screening Panel (24) (Cat# 333812) 3 μL of each library was pooled together into one tube. The pooled library concentration was measured in ng/μL using the Qubit DNA HS assay. Quality of the pooled library was checked using a Bioanalyzer. Size was measured at 636bp. The pooled library was diluted to 10 nM using 10 mM Tris-HCl pH 8.5 0.1% Tween 20 buffer. Sequencing was done on a Nano-300 cycle test chip (MS-103-1001) to confirm coverage of all libraries. Libraries with low reads were spiked in at higher concentrations to increase coverage. The re-pooled library was then ran on the MiSeq using an Illumina MiSeq Reagents kit v3 (600 cycles) (MS-102-3003) at 10 pM set at the below parameters according to kit instructions: 16s rRNA sequencing
GEO Accession
GSM5034309