title: GSM8377829: DA3K1; Mesocricetus auratus; RNA-Seq
library_construction_protocol: Extraction of RNA was performed using the AllPrep DNA/RNA Mini Kit (Qiagen). In short, hamster kidney tissues were homogenized in 350 μL of RLT buffer and then applied to a DNA spin column. The homogenized lysate was centrifuged to extract DNA, and the resulting supernatant containing RNA and protein was collected. Next, absolute ethanol was added, gently mixed with a pipette, transferred to an RNeasy® spin column, and centrifuged to remove proteins from the mixture. The RNA portion was washed with 700 μL of RW1 buffer and 500 μL of RPE buffer, respectively. Finally, the RNA was eluted with 50 μL of RNase-free water and stored at -80°C until use. (1) mRNA was isolated by Oligo(dT)-attached magnetic beads.  (2) mRNA was then randomly fragmented in fragmentation buffer.  (3) First-strand cDNA was synthesized with fragmented mRNA as template and random hexamers as primers, followed by second-strand synthesis with addition of PCR buffer, dNTPs, RNase H and DNA polymerase I. Purification of cDNA was processed with AMPure XP beads.  (4) Double-strand cDNA was subjected to end repair. Adenosine was added to the end and ligated to adapters. AMPure XP beads were applied here to select fragments within size range of 300-400 bp.  (5) cDNA library was obtained by certain rounds of PCR on cDNA fragments generated from step (4).