GSM6797146: GC-JC-8721-1; Dictyostelium discoideum; RNA-Seq
Cell suspensions were loaded to the 10X Chromium Single Cell A Chip (PN-1000009) using the Chromium 3' Library and Gel Bead Kit v2 (PN-120267) as described by the manufacturers (10X Genomics, California). 14 cycles of cDNA amplification were performed on the purified GEM-RT product, and cDNA was examined for quality using the Agilent 2200 Tapestation with the High-sensitivity D5000 screentape and reagents (Agilent Technologies, Waldbronn, Germany), and the Qubit 2.0 Fluorometer and Qubit dsDNA HS Assay Kit (Life Technologies, California, USA). 35 µL of cDNA was used to prepare the 10x 3'RNA library and 12 and 13 cycles were used for sample index PCR of replicates 1 and 2 respectively. Final cleaned libraries were quantified using the Qubit 2.0 Fluorometer and Qubit dsDNA HS Assay Kit and average fragment size checked using the Agilent D1000 screentape and reagents. The final library was run on a NextSeq500 Mid-output 150-cycle kit with a 26[8]98 cycle configuration to generate 130 million read pairs in total.