title: GSM7504822: Transplant Patient Blood B cell Memory; Homo sapiens; OTHER
library_construction_protocol: B cell subsets were sorted directly into lysis buffer (Norgen Single Cell RNA Purification Kit, Cat #51800) with 1% mercaptoethanol, lysates were snap frozen on dry ice and stored at -80°C. Total cellular RNA was extracted from bulk sorted B cell and ASC subsets according to the manufacturer's protocol. cDNA preparation was performed (BioRad iScipt, Cat #1708891) per manufacturer's instructions, and DNA amplicon products where generated using Ig-specific variable and constant gene primers. Samples were indexed (Nextera Index Kit, Illumina), purified, quantitated (KAPPA Library Quantitation Kit, Roche) and pooled into libraries. Following generation, libraries were denatured per manufacturer instructions (Illumina) and loaded onto a 600-cycle V3 MiSEQ cartridge (Illumina) for sequencing.