GSM2944301
GSM2944301: K8W1A1; Homo sapiens; RNA-Seq
A modified Smart-seq2 protocol was applied for single-cell RNA-seq. Briefly,  single cell was picked into the lysis buffer by mouth pippete. The reverse transcription reaction was performed with a 24 oligo (dT) primer, which was  anchored with a 8 bp cell specific barcode and a 8 bp unique molecular identifiers (UMIs). After the first-strand reaction, the cDNA was amplified by 19 cycles. Then the amplified cDNA of the cells of the same fetal sample were pooled together to proceed the following steps. Biotinylated pre-indexed primer was used to amplify the PCR product by additional 4 cycles to introduce biotin tags to the 3' end of the amplified cDNA. About 300 ng DNA was sheared to around 300 bp by Covaris S2 and the 3' ternimal of the DNA was captured by Dynabeads® MyOne™ Streptavidin C1 beads (Thermo Fisher). RNA seq library was constructed using Kapa Hyper Prep Kit (Kapabiosystems) and subjected to 150 bp paired-end sequencing on illumina HiSeq 4000 platform (sequenced by Novogene).
GEO
GSM2944301
GEO Accession
GSM2944301