CORPRO-Seq of mus musculus: cecal contents
DNA was extracted from intestinal samples using mechanical disruption (bead-beating with 0.1 mm zirconium beads) in phenol:chloroform:IAA in the same manner as described by McNulty et al. in a previous publication (PMID: 22030749).Libraries were prepared according to a slightly modified version of the protocol accompanying the Illumina Genomic DNA Sample Prep Kit. Briefly, total bacterial gDNA was sonicated in a BioRuptor XL water bath sonicator, cleaned up and concentrated using a Macherey-Nagel PCR Purification column. DNA was blunted and poly-A tailed. The A-tailed molecules were ligated to the relevant barcoded Illumina adapter sequence. Adaptered-DNA was then size-selected by agarose gel electrophoresis. Fragments of the appropriate size were PCR amplified and purified, after which the purified PCR products were give to the University of Wisconsin-Madison Biotechnology Center for quality assurance and library sequencing following the manufacturer's protocols. Organisms used: Blautia hansenii ATCC 27752, Dorea longicatena DSM 13814, Marvinbryantii formatexigens DSM 14469, Bacteroides caccae ATCC 43185, Bacteroides thetaiotaomicron VPI-5482, Providencia stuartii ATCC 29914, and Collinsella aerofaciens ATCC 25986.
SUB4467481
CIIH8