title: GSM4830961: mmB FeLO D17-2; Homo sapiens; RNA-Seq
library_construction_protocol: Tissues were harvested and lysed by adding 500μL Trizol (Life Technologies) directly to the tissue culture well and storing at -80°C.  For extraction, lysate was thawed on ice, 100μL chloroform was added, vortexed for 30 seconds, and centrifuged at 12,000g for 15 minutes at 4°C.  After centrifugation the aqueous phase was transferred to a QIAGEN gDNA eliminator column and centrifuged at 10,000g for 1 minute at room temperature. The flow-through was mixed with an equal volume of 70% EtOH and transferred to an RNEasy mini spin column.  The manufacturer protocol for the RNEasy Plus Mini Kit (QIAGEN) was followed for the rest of the extraction procedure. Libraries for RNA-Seq were prepared with KAPA Hyper Stranded RNA-Seq Kit. The workflow consists of mRNA enrichment, cDNA generation, and end repair to generate blunt ends, A-tailing, adaptor ligation and PCR amplification. Different adaptors were used for multiplexing samples in one lane.