GSM2936905
GSM2936905: RNA_FAP3_N_B1; Homo sapiens; RNA-Seq
Single cells were picked into scRNA-seq lysis buffer or into scMALBAC 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. And the cells in scMALBAC lysis buffer were centrifuge for 1min at 7,000 rpm and then incubate at 50℃ for 3 hours to release genomic DNA from nucleosome. The remaining cells were used to extract genomic DNA 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 sequencing on Illumina 4000.  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 Prep Kits with PCR Library Amplification/Illumina series (KAPA, KK8054) and sequencing on Illumina 4000. single cell RNA-Seq; Single cell whole genomic sequencing
GEO
GSM2936905
GEO Accession
GSM2936905