title: RNA-Seq of control
design: Illumina sequencing was performed at Beijing GenomicsInstitute (BGI)-Shenzhen, China. mRNA with poly(A) tail was isolated from 20 mg total RNA treated with DNase I using Magnetic Oligo (dT) Beads (Illumina). The mRNA was fragment- ed into small pieces (200700 bp) by treatment with divalent cations at 94uC for 5 minutes. With random hex-amer primers (Illumina), the double-stranded cDNA was synthesized using the SuperScript double-stranded cDNA synthesis kit (Invitrogen) and was further subjected to end-repair using T4 DNA polymerase, the Klenow fragment, and T4 polynucleotide kinase followed by a single A base addition using Klenow 39 to 59 exo-polymerase, then was ligated with an adapter or index adapter using T4 DNA ligase. To select the proper templates for downstream enrichment, the products of ligation reaction were purified on 2% agarose gel. The cDNA fragments (about 200 bp) were recovered from the gel. Fifteen rounds ofPCR amplification were carried out to enrich the purified cDNA template using PCR primer PE 1.0 and 2.0 (Illumina) with Phusion DNA polymerase. Finally, the cDNA library was constructed with 200 bp insertion fragments. After validating on an Agilent Technologies 2100 Bioanalyzer, the library was sequenced using Illumina HiSeqTM 2000 according to the manufacturers instruction.Data