title: GSM4874279: Normal-conditioned_2; Bos taurus; RNA-Seq
library_construction_protocol: Frozen-thawed liver samples (20 mg) were finely chopped with scalpel blades (RNA away sprayed) and homogenized in 1 mL of Purezol (Qiagen, Antwerp, Belgium). Total RNA was isolated using the Aurum Total RNA Mini Kit (BioRad Laboratories Inc., Hercules, CA) according to the manufacturer's instructions. After RNA isolation, a DNase treatment with 6 μL of RQ1 RNase-Free DNase (1 U/μL, Promega, Leiden, the Netherlands) and 3 μL of RQ1 DNase 10x Reaction Buffer (Promega) was performed for 15 min at room temperature. Then, RNA was purified by spin-column centrifugation (purification step: 2 times for 7 min at 16,100 × g at 20°C, concentration step: 1 time for 3 min at 1,000 × g at 20°C; Amicon Ultra-0.5 centrifugal filters, Merck Millipore, Billerica, MA). Quantity and purity of RNA were evaluated with the Nanodrop ND-1000 spectrophotometer (Nanodrop Products, Thermo Fisher Scientific, Wilmington, DE). Additionally, RNA integrity was verified by visualization of the ribosomal bands on a 2% agarose gel. Genomic DNA contamination was verified by a minus reverse transcriptase control PCR with LRP10 primers. All RNA samples had a purity between 1.91 and 2.16 (optical density 260/280 ratio), clear ribosomal RNA bands on gel, and were free of genomic DNA. Total RNA was prepared and sent to NXTGNT. The sequencing library was constructed using the QuantSeq 3' mRNA-Seq Library Prep Kit FWD for Illumina from Lexogen and was sequenced as SE75 on an Illumina Nextseq 500
attributes.tag: GEO Accession
attributes.value: GSM4874279