Dual RNA sequencing of the host transcriptome and metatranscriptome at multiple intestinal sites of common marmosets

This project investigated the host-microbiome interaction within multiple intestinal sites of the non-human primate species, common marmosets. Dual RNA sequencing was performed at three intestinal sites, the cecum, transverse colon, and rectum of five marmosets. After the gastrointestinal tract was excised from each marmoset, the samples from each site were immediately frozen in liquid nitrogen and preserved at -80 °C. Within a week of dissection, these samples were crushed and homogenized in solution D, which contains guanidinium for ribonuclease inhibition, to protect against degradation. RNA extraction was achieved using a combination of the acid-guanidinium-phenol-chloroform method and bead crushing. Ribosomal RNA (rRNA) was depleted with the Ribo-Zero Plus rRNA Depletion Kit (Epidemiology) from Illumina. Sequencing libraries were prepared using Illumina's TruSeq Stranded Total RNA HT Kit and the sequencing was performed with Illumina's NovaSeq 6000 v1.5.