Poly(A) RNA-seq Library Preparation

The polyadenylated fraction of RNA isolated from eight samples was used for 100bp paired-end RNA-seq with coverage 43.8 ± 4.1 sd million pairs read per sample. We used the SMARTer^® Ultra™ Low RNA Kit for Illumina^® Sequencing (Clontech) followed by the NEBNext^® DNA Library Prep Master Mix Set for Illumina^® to contruct poly(A) selected pair-end sequencing libraries. Both kits were used as per the manufacturer’s instructions except that published in-house custom indexes were used (45 (link)). The resulting multiplexed libraries were sequenced using Illumina TruSeq v3 chemistry. After indexing, all samples were combined into a single library and sequenced on two lanes of an Illumina HiSeq 2000 System.