M.CH.DxB
Library preparation was performed as recommended by the manufacturer (Illumina, Hayward, CA). Briefly, total RNA was extracted using the RNeasy Mini kit with DNase treatment (Qiagen, Valencia, CA). Poly(A) mRNA was isolated and fragmented, and first-strand cDNA was prepared using random hexamers. Following second-strand cDNA synthesis, end repair, addition of a single A base, adaptor ligation, and agarose gel isolation of ∼200-bp cDNA, PCR amplification of the ∼200 bp cDNA was performed. Liver samples were sequenced using the Illumina GAIIX sequencer to a coverage of ∼40 million single-end reads of 75 bp.