Analysis of Akkermansia muciniphila in Mulberry Galacto-Oligosaccharide Medium by Comparative Transcriptomics
An RNA-seq transcriptome library was prepared with the TruSeq RNA sample preparation Kit from Illumina (San Diego, CA, USA) using 2 g of total RNA. Shortly after preparation, ribosomal (r)RNA depletion, instead of poly (A) purification, was performed using the Ribo-Zero Magnetic kit (Epicenter), then all mRNAs were broken into short fragments by adding fragmentation buffer. Double-stranded cDNA was synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen) with random hexamer primers (Illumina). The ligation products were size-selected via agarose gel electrophoresis, amplified via PCR using Phusion DNA polymerase (NEB) for 15 cycles, and sequenced using Illumina HiSeqTEN (2  150 bp read length).