The ncRNA deletion strategy and composition of the ncRNA deletion collection. (A) A schematic of the PCR-based ncRNA deletion strategy. The kanMX4 ncRNA deletion cassette was amplified from the pFA6a-kanMX4 plasmid using primers “Up primer (101mer)” and “Down primer (101mer).” The primers contain two barcodes (Tag), unique for each ncRNA and genome target homologous region (HR) sequences (upstream of and downstream from the ncRNA) to allow ncRNA disruption by homologous recombination. The kanMX4 ncRNA deletion cassettes were integrated into the genome at least 200 bp away from the closest open reading frame (ORF) start codon. (B) Composition of the ncRNA deletion collection. The “other ncRNA” are NME1, RPR1, RUF21, and TLC1.
