GSM4160345: mx0019; Homo sapiens; RNA-Seq
4 mm skin biopsies of skin from atopic dermatitis patients or deidentified healthy margin surgical tissue were obtained; skin samples were placed in RNA Later and tissue was lysed using a bead homogenizer followed by RNA isolation using the RNeasy mini kit (QIAGEN) rRNA was depleted using RiboZERO kit (Illumina), mRNA was fragmented in buffer containing 40 mM Tris acetate (pH 8.2), 100 mM potassium acetate, and 30 mM magnesium acetate and heated to 94C for 150 s. mRNA was reverse transcribed to yield cDNA using SuperScript III RT enzyme (Invitrogen, per manufacturer's instructions) and random hexamers. A second strand reaction was performed to yield dscDNA. cDNA was blunt ended, had an A base added to the 30 ends, and then had Illumina sequencing adapters ligated to the ends. Ligated fragments were then amplified for 12 cycles using primers incorporating unique index tags. Fragments were sequenced on an Illumina HiSeq-3000 using single reads extending 50 bases.
GEO Accession
GSM4160345