title: GSM4658686: Fish1; Salmo salar; RNA-Seq
library_construction_protocol: Lens tissue was thoroughly homogenized before RNA extraction using a Precellys 24 homogenizer and ceramic beads CK28 (Bertin Technologies, Montigny-le-Bretonneux, France). Total RNA was extracted using the BioRobot EZ1 and RNA Tissue Mini Kit (Qiagen, Hilden, Germany), treated with DNase. The RNA integrity number (RIN) of RNA extracted for RNA-seq (2N: n=12, 3N: n=14) was 8.2±0.1 (n=26) (mean ± SEM). according to the manufacturer's instructions and eluted in 50 μL RNase-free MilliQ H2O. Poly (A) mRNA was isolated using magnetic beads with oligo (dT) from total RNA obtained from the lens samples. Fragmentation buffer was added to shred mRNA to short reads. Using these short fragments (about 200 bp) as templates, random hexamer primers were applied to synthesize first-strand cDNA. Second-strand cDNA was synthesized using buffer, dNTPs, RNaseH, and DNA polymerase I. QiaQuick PCR extraction kit (Qiagen) was used to purify short double-stranded cDNA fragments according to manufacturer's instructions. These fragments were then resolved with EB buffer for end reparation, added poly (A), and ligated to the sequencing adapters. After agarose gel electrophoresis, the suitable fragments were selected for PCR amplification as templates.
attributes.tag: GEO Accession
attributes.value: GSM4658686