title: RNA-Seq of estrogen treated liver: 10 weeks old female chicken
design: Total RNA was extracted from the chicken liver tissues using TRIzol  reagents following the manufacturer s manual (Invitrogen, Carlsbad, CA). Qualified total RNA was further purified by RNA Clean XP Kit (Cat#A63987, Beckman Coulter, Inc. Kraemer Boulevard Brea, CA, and USA) and RNase-Free DNase Set (Cat#79254, QIAGEN, GmBH, Germany). Libraries were constructed using the TruSeq Stranded Total RNA LT -(with Ribo-Zero TM Gold)-Set B (Cat#RS-122-2302, Illumina, San Diego, CA) according to the manufacturer s protocol. Briefly, the mRNA was isolated from the total RNA using oligo (dT) beads with two rounds of oligo-dT purification, followed by fragmentation of RNA into short mRNA fragments (each of approximately 360bp). Then the cleaved RNA fragments were synthesized into single-strand cDNA using superscript II reverse transcriptase (Cat#18064014, Invitrogen) and random hexa-primers. The second-strand was synthesized using DNA polymerase I (Invitrogen). After end repair and A-tailing, the synthesized double stranded cDNA fragments were amplified by PCR and amplification products were purified before sequencing.