GSM3823867: DEC.70818; Homo sapiens; RNA-Seq
Tissue samples (5-15 mg) were stored with 250 μl RNAlater RNA Stabilization Reagent (QIAGEN, Hilden, Germany) at -80°C in the CSMC Prenatal Repository until further processing. Briefly, tissue was thawed on ice with 600 μl of RLT Plus lysis buffer (QIAGEN) and 1% β-mercaptoethanol added to each sample. Tissue was homogenized by passing through increasingly thinner single-use needles (22G, 25G, 27G) attached to a 1 mL sterile, RNase-free syringe. The homogenates were loaded onto AllPrep spin columns and the remainder of the protocol was performed following manufacturer instructions for the AllPrep DNA/RNA Mini Kit (QIAGEN). The RNA elution was used for total RNA-sequencing. RNA-seq libraries were constructed using Illumina TruSeq Stranded Total RNA LT with Ribo-zero kits (Illumina, San Diego).