GSM5515747: DHA4_STD; Homo sapiens; RNA-Seq
Fat layer containing mammary epithelial cell contents from fresh milk sample placed into a 2mL tube with 500 μL of TRIzol LS (Thermo Fisher Scientific, Waltham, MA) To extract the mRNA, 200 μL of chloroform was added and samples were gently inverted by hand for 15 seconds and incubated for 5 minutes at room temperature. The pure mRNA was isolated and purified using the PROMEGA Maxwell 16™ integrated system (Promega Corporation, Madison, WI) The samples were checked for quality and quantity using the Nanodrop 1000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA) and the Agilent 2100 Bioanalyzer (Agilent, Santa Clara, CA). Samples sequenced using the Illumina HiSeq 2000 (Illumina, Inc., San Diego, CA) and reads were trimmed for quality and aligned using BaseSpace RNA-seq Alignment v2.02. (Illumina, Inc., San Diego, CA) and STAR (Illumina, Inc., San Diego, CA). Salmon (Illumina, Inc., San Diego, CA) was used for quantification (TPM). Standard poly-A RNA-seq SR 1x85bp, 50M reads