ena-STUDY-Cell and Developmental Biology-28-07-2021-11:58:10:569-664
D. discoideum Cdl1a knockout - wild-type RNAseq comparison
D.discoideum wild-type Ax2 and cdl1a (DDB_G0286351) knockout cells were developed at 22ºC on non-nutrient agar into mid to late fruiting bodies. Total RNA was isolated from three independent experiments and enriched for mRNA using poly-T–linked magnetic beads. Barcoded cDNA libraries were constructed using NEBNext Ultra II Directional RNA Kit for Illumina, following manufacturer's instructions, and checked for quality using the Agilent TapeStation DNA D1000 HS Kit. The six bar-coded libraries were normalized to 10 nM and combined into one pool, which was sequenced as paired-end 75-bp reads at ~16 million reads per sample using the Illumina mid-output NextSeq 500 platform.
D.discoideum wild-type Ax2 and cdl1a (DDB_G0286351) knockout cells were developed at 22ºC on non-nutrient agar into mid to late fruiting bodies. Total RNA was isolated from three independent experiments and enriched for mRNA using poly-T–linked magnetic beads. Barcoded cDNA libraries were constructed using NEBNext Ultra II Directional RNA Kit for Illumina, following manufacturer's instructions, and checked for quality using the Agilent TapeStation DNA D1000 HS Kit. The six bar-coded libraries were normalized to 10 nM and combined into one pool, which was sequenced as paired-end 75-bp reads at ~16 million reads per sample using the Illumina mid-output NextSeq 500 platform.