GSM7287632: Patient 23, mRNA-derived cDNA; Homo sapiens; RNA-Seq
After enzymatic dissociation of the embryo; CFSE+ hCD45+ Live cells were sorted by flow cytometry Libraries were prepared using Chromium Single Cell 5ʹ v1.1 protocol (10x Genomics). mRNA libraries were prepared according to the manufacturer's instructions HTO libraries were prepared according to the manufacturer's instructions. For each sample, the three conditions (before and after graft with RCHOP or vehicle treatment) were done and pooled in the same experiment with the TotalSeq -C listed above BCR libraries were amplified accoring to 10X Genomics guidelines. 800 pg of amplified sequences was used with Nextera XT reagents (Illumina San Diego, California) to construct the final BCR library of each sample by PCR using i5 forward and Nextera XT i7 reverse primers. The three libraries were quantified using the Library Quantification Sample Kit Kapa (Illumina-Uni San Diego, California) Following the final bead purification, all three libraries were pooled as 80% mRNA library, 10% HTO library and 10% BCR library before sequencing.