title: GSM4162517: IDH1_H2O2_2: bulk RNA seq; Homo sapiens; RNA-Seq
library_construction_protocol: Single-cell RNA-seq: Single-cell RNA-seq libraries were constructed according to the STRT-seq protocol with modifications, as previously described. Briefly, individual oocytes or somatic cells were picked into lysis buffer by mouth pipetting. Then, the mRNA in lysates was reverse transcribed with SuperScript II reverse transcriptase (Thermo Fisher Scientific, 18064-071), and an 8 nt cell-specific barcode was added to the cDNA from each cell. Absolute counting of molecules was performed with the 8 nt unique molecular identifiers (UMIs). The synthesized cDNAs from cells with different barcodes were pooled together and amplified with 19-21 (somatic cells) or 13 (oocytes) cycles, followed by fragmentation with Covaris S2 (Thermo Fisher) and enrichment of fragments of interest via Dynabeads MyOne Streptavidin C1 beads (Thermo Fisher, 65001). The enriched fragments were constructed into libraries using a KAPA Hyper Prep Kit (KAPA, KK8504) and sequenced on an Illumina HiSeq 4000 platform with a 150-bp paired-end read length by Novogene. Bulk RNA-seq: Total RNA was extracted from one million cells using TRIzol reagent (Invitrogen). 3 ug per sample were constructed into libraries through NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, E7530L) following manufacturer's manuals. Sequencing data were generated via on Illumina HiSeq 4000 platform with 150-bp paired-end read length by Novogene.
attributes.tag: GEO Accession
attributes.value: GSM4162517