title: GSM3928109: Mammary epithelial cells, Control2; Bos taurus; RNA-Seq
library_construction_protocol: Total RNA was isolated using the Magen HiPure Total RNA Mini Kit (Magen, Guangzhou, China). RNA concentration was measured using a NanoDrop 2000 (Thermo Scientific, Wilmington, DE) and RNA quality was assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). All samples used in this study had an RNA integrity number > 8 (Supplementary Table S2). Six individual libraries were constructed: three from high-milk-fat samples and three from control samples. RNA-seq transcriptome libraries were prepared following the TruSeq RNA Sample Preparation Kit (Illumina, San Diego, CA) using 5 μg of total RNA. mRNA was isolated according to standard poly(A) selection methods using oligo (dT) beads and then fragmented in fragmentation buffer. cDNA was synthesized using a SuperScript Double-stranded cDNA Synthesis Kit (Invitrogen) with random hexamer primers (Illumina). Synthesized cDNA was subjected to end repair, phosphorylation, and A-base addition. Libraries were size-selected for cDNA target fragments of ~200-300 bp on 2% low-range ultra agarose and then amplified followed by 15 cycles of PCR. After quantification on a TBS380 fluorometer (American Instrument Exchange, Inc., Haverhill, MA), paired-end RNA-seq sequencing libraries were sequenced on an Illumina HiSeq 4000 (2 × 150-bp read length)
identifiers.namespace: GEO
identifiers.id: GSM3928109