GSM2976295: P5-LyM1-2-lyM1-3; Homo sapiens; RNA-Seq
Single cells were picked into scRNA-seq lysis buffer by using mouth pipette. Then the cells in scRNA-seq lysis buffer were vortexed for 45 second and heated at 72℃ for 3min to release RNA. Bulk genomic DNA were extracted according to the manufacturer protocol of DNeasy Blood & Tissue Kit (QIAGEN). Modified STRT-Seq protocol: we modified the STRT-Seq protocol for application to a large number of cells. Briefly, the 8bp barcodes was used to label individual cells. So the cDNA of cells with different barcodes can be pooled together for library constructions. And we also enrich the 3' cDNA by the affinity interaction between biotin and Dynabeads® MyOne™ Streptavidin C1 beads. Then the fragmented DNA inserts were library constructed according to the manufacturer procedures of KAPA Hyper Prep Kits with PCR Library Amplification/Illumina series (KAPA, KK8054) and sequenced on Illumina 4000. Bulk genomic DNA library construction protocol: The extracted genomic DNA was fragmented by sonication into 300bp and then library constructed according to the manufacturer procedures of KAPA Hyper Pre.
GEO
GSM2976295