GSM8326146: AM1_2; Homo sapiens; RNA-Seq
The cellular pellet samples were used to isolate total RNA with TRI Reagent (Merck, USA) and RapidOutput DNA removal kit (ThermoFisher Scientific, USA). A Qubit 4 fluorometer (ThermoFisher Scientific, USA) and Fragment Analyzer (Agilent, USA) were used to analyse the isolated RNA for quantity and quality, respectively. Complementary DNA (cDNA) libraries were synthesized using the NEBNext® Poly(A) mRNA Magnetic Isolation Module and NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (NEB, UK). The concentration and quality of cDNA libraries were assessed using a Qubit 4 fluorometer (ThermoFisher Scientific, USA) and Fragment Analyzer (Agilent, USA). The cDNA samples were then diluted to 4 nM and sequenced on the Illumina NextSeq® platform with NextSeq® 500/550 High Output Kit v2.5 (150 cycles) (Illumina, USA), generating 75 bp pair-end reads, and on the Illumina NovaSeq® 6000 platform with NovaSeq® S4 300 kit (Illumina, USA), generating 150 bp pair-end reads.