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Fifty ng of the 5'-end scRNA-seq cDNAs derived from rhesus macaque primary neuronal cells cultured at day 15 was amplified for six cycles with the high-fidelity 2× PCR Master Mix (New England Biolabs, Cat. No. M0541), using cDNA amplification primers provided in the 5'-end scRNA-seq kit (10x genomics, Cat. No. PN-1000265). The amplified cDNAs were then purified and subjected to size selection using BluePippin size selection system (Sage science, BluePippin) with 1.5% gel (Sage science, Cat. No. BDF1510) to collect desired cDNAs with fragment size > 1,900 bp. The size-selected cDNAs were further amplified for 5 cycles and then were applied to PacBio sequencing library construction using SMRTbellTM Template Prep Kit (PacBio, Cat. No. 100-938-900) according to the instructions of the manufacturer. |
ISO-Seq |
TRANSCRIPTOMIC SINGLE CELL |
PolyA |
SINGLE
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