GSM7412338: PE_tissue10X; Homo sapiens; RNA-Seq
Graft tissue generated by skeletal stem cells was mechanically and enzymatically digested, stained with antibodies, and living Venus-expressing populations purified via FACS. cDNA was prepared according to 10X Genomics manufacturer instructions for 3' v3.1. 25% (10ul) of cDNA was used for standard 10X Genomics library preparation protocol. Another 25% was used for clonal barcode amplification. The extra 10 µL of cDNA were amplified using a combination of Partial Read 1 primer  (5'-CTACACGACGCTCTTCCGATCT-3') and an adaptor-FBP1 (Read 2 primer + barcode recognition site) primer (5'-GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTCCGACCACCGAACGCAACGCACGCA-3'). Barcode amplification was run as recommended for cDNA reaction (10X protocol step 2.2) plus 4 extra cycles. From there the product was handled as per standard protocol starting from the index primer reaction without any fragmentation or ligation steps.