GSM7676907: Donor 4 Hiltonol+pRNA stimulated BDCA3; Homo sapiens; RNA-Seq
RNA was harvested using Trizol (Invitrogen, MA, USA), following the standard protocol. Quality control of the isolated RNA (concentration, RIN, 28S/18S and size) was performed with Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, USA). After extraction of total RNA and treatment with with DNase I, Oligo(dT) are used to isolate mRNA. Mixed with the fragmentation buffer, the mRNA are fragmented. ThencDNAis synthesized using the mRNA fragments as templates. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. The suitable fragments are selected for thePCRamplification. During the QC steps, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-TimePCRSystem are used in quantification and qualification of the sample library. Then the library is sequenced using Illumina HiSeq 4000 or other sequencer when necessary.