RNA-seq of human colonocytes cultured with gorilla microbiome
Poly-adenylated mRNA was isolated from thawed cell lysates using the Dynabeads mRNA Direct Kit (Ambion) following the manufacturers instructions. RNA-seq libraries were prepared using a protocol modified from the NEBNext Ultradirectional (NEB) library preparation protocol to use Barcodes from BIOO Scientific added by ligation, as described in Richards et al. (Richards et al., 2019). The libraries were then pooled and sequenced on two lanes of the Illumina Next-seq 500 in the Luca/Pique-Regi laboratory using the high output kits for 75 cycles to obtain paired-end reads.