GSM7143068: DU4475 - suspension, matrigel, soft fibrin, stiff fibrin; Homo sapiens; RNA-Seq
Total RNA was extracted using RNeasy Mini kit (Qiagen) and purified with RNA Clean and Concentrator kit together with DNAse treatment (Zymo). The library was contructed using the bulk RNA barcoding (BRB) sequencing method. Total RNA (10 ng) was barcoded, converted to cDNA, and amplified by PCR. The PCR products were pooled together, purified with 0.6X Agencourt AMPure XP Beads (Beckman Coulter) and tagmented using the Nextera kit (Illumina). Library quantity was assessed using a Qubit 2 fluorometer (Invitrogen) and the Qubit DNA HS Assay Kit (ThermoFisher Scientific) and library quality using the LabChip GXII Touch HT (PerkinElmer), with the DNA High Sensitivity Assay (PerkinElmer) and the DNA 5 K / RNA / Charge Variant Assay LabChip (PerkinElmer). The libraries were then sequenced using Illumina NextSeq 500 (Illumina). 3' RNA-Seq (Drop-seq-like)