SC DNA-Seq of immortalized Lbeta2 gonadotrope cells
Single cells were picked into 2.5 l of PBS using the CellRaft (Cell Microsystems) SC picking system, following the manufacturers guidelines. DNA amplification was performed using the Rubicon genomics PicoPLEX WGA Kit (Cat # R30050) with the adjustment of final amplification cycles to 8, following the manufacturers instructions. Purification was carried out using AMPure beads at a 0.9X concentration. Following amplification, 300 ng of DNA was used to create Ion Torrent libraries using the NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent (Cat # E6285L) with a few minor modifications. The adapter ligation was completed with 3 l of NEXTflex DNA Barcodes (cat #NOVA-401004) and the final amplification step was omitted. After the libraries were purified using AMPure beads, 250-bp fragments were size-selected with the Invitrogen E-gel size selection system. The libraries were sequenced at an average of 0.2X coverage on the Ion Proton.