title: GSM2898235: tlr_46hr_b3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated using a RNeasy plus Micro RNA kit (Qiagen, Hilden, Germany) [Trombetta et al CPMB 2014] [Whole Transcriptome Amplification]: Two μL of extracted RNA were added to 8 μL of water and cleaned with 2.2x volume beads. 1 μL of this cleaned RNA was used in a WTA reaction. Whole transcriptome amplification was performed on these samples analogous to the amplification of the single cell samples. [Preparation of cDNA Libraries  for RNAseq]: WTA products were diluted to a concentration of 0.1 to 0.4 ng/μL and tagmented and amplified using Nextera XT DNA Sample preparation reagents (Illumina). Tagmentation was performed according to manufacturer's instructions, modified to use ¼ the recommended volume of reagents, extended tagmentation time to 10 minutes and extended PCR time to 60s. PCR primers were ordered form Integrated DNA Technologies. Nextera products were then cleaned twice with at 0.9x volume of SPRI beads and eluted in water. The library was quantified using Qubit and analyzed using a high sensitivity DNA chip. The library was diluted to 2.2 pM and sequenced on a NextSeq 500 (Illumina)
attributes.tag: GEO Accession
attributes.value: GSM2898235