Total RNA was extracted by using of RecoverAll Total Nucleic Acid Isolation Kit (Ambion). Poly(A)+RNA was selected and enriched with oligo(dT) using the Oligotex mRNA mini Kit (Qiagen).  The Poly(A) RNA was heat sheared to 100~200nt fragments. The sheared RNA was then subjected to first and second strand cDNA synthesis, followed by additional steps of library construction as described previously. The libraries were sent to the Stanford Genomic Center to be sequenced for the first 36bp at the 5'end of mRNA fragments with Illumina machines.
GEO Accession
GSM1447368