GSM1419093: WRR; Gallus gallus; ncRNA-Seq
Total RNA of 8 breast muscles was isolated using Trizol Reagent (Invitrogen, Carlsbad, CA, USA) according to the protocol of manufacture. RNA libraries were prepared for sequencing using standard Illumina protocols. Briefly, total RNA from 4 birds within each group was mixed in equal amounts to generate a pooled sample using Agilent 2000. Ribosomal RNA was removed from total RNA. Subsequently, the RNA was sonicated to produce RNA fragments ranging from 200-500 nt. The RNA fragments were used to generate the first strand cDNA synthesis with normaldNTPs. Then, the second strand cDNA synthesis was obtained with dTTP changed to dUTP. After purifying with QIAquick Gel Extraction Kit (Qiagen, Valencia, CA, USA), the addition of EB buffer solution, end repairing, the addition of single “A” base to 3’ terminal and Y-adaptor ligation, and the degradation of the second strand cDNA synthesis by Uracil-N-Glycosylase enzyme, qualified RNA was excised from the gel by fragment size selection and then used for PCR amplification. Following library qualification, RNA libraries were sequenced on the Illumina Hiseq 2000 (Illumina, San Diego, CA, USA)
GEO Accession
GSM1419093