title: GSM2938940: CMP_sc8; Mus musculus; RNA-Seq
library_construction_protocol: RNA libraries were prepared for sequencing using a modified Smart-seq2 protocols. Each single LT-HSC or CMP was placed into the lysis buffer by mouth pipette,The reverse transcription reaction was performed with 25 nt oligo(dT) primer anchored with an 8 nt cell-specific barcode and 8 nt unique molecular identifiers (UMIs).After the first-strand synthesis, the second-strand cDNAs were synthesized, and the cDNAs were amplified by 17 cycles of PCR. The amplified cDNAs of the single cells were then pooled together for the following steps. Biotinylated pre-indexed primers were used to further amplify the PCR product by an additional 4 cycles of PCR to introduce biotin tags to the 30 ends of the amplified cDNAs. Approximately 300 ng cDNA was sheared to approximately 300 bp by Covaris S2, and the 30 terminal of the cDNA was captured by Dynabeads MyOne Streptavidin C1 beads. The RNA-seq library was constructed using a Kapa Hyper Prep Kit and subjected to 150 bp paired-end sequencing on an Illumina HiSeq 4000 platform. Single cell RNA-seq
attributes.tag: GEO Accession
attributes.value: GSM2938940