Differential gene expression profiling of the goose pineal organ |
Total RNA was extracted from each pineal organ using TRIzol reagent (Invitrogen, USA) according to the manufacturer’s instructions. RNA quality was confirmed on 1% agarose gels. RNA purity and concentration was determined using a NanoPhotometer® spectrophotometer (IMPLEN, CA, USA) and Qubit® RNA Assay Kit (Qubit® 2.0 Flourometer; Life Technologies, CA, USA) according to the manufacturer’s instructions. RNA integrity was assessed using the RNA 6000 Nano Assay Kit (Bioanalyzer 2100 system; Agilent Technologies, CA, USA). Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit (Illumina®, NEB, USA) and following the manufacturer’s recommendations. Clustering of the index-coded samples was performed on a cBot Cluster Generation System using the TruSeq PE Cluster Kit v3-cBot-HS (Illumina) according to the manufacturer’s instructions. After cluster generation, the library preparations were sequenced on an Illumina HiSeq platform and 125 bp/150 bp paired-end reads were generated. |
RNA-Seq |
GENOMIC |
PolyA |
SINGLE
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