title: Shotgun metatranscriptomic sequencing of infant fecal material
design: Total RNA was extracted from fecal samples using the AllPrep PowerFecal DNA/RNA kit (Qiagen) and subsequently treated with DNase. Purified RNA quality and concentration were measured using the Fragment Analyzer (Agilent). Illumina sequencing libraries were constructed with the ScriptSeq Complete Gold Kit (Illumina) without performing the rRNA removal step, resulting in library molecules with an average insert size of 150 bp. Sequencing was performed on Illumina's NextSeq platform in a paired end 75 bp configuration, resulting in an average of 54 million paired end reads per sample.