WT1
Total RNA extracts were rRNA depleted and RNAseq libraries prepared, using a QIAseq FastSelect rRNA 5S/16S/23S kit (QIAGEN) and NEBNext Ultra II Directional RNA Library Prep kit (New England Biolabs). Individual libraries were indexed using Index Primers from NEBnext Multiplex Oligos for Illumina Sets 1 and 2 (New England Biolabs); quantified using a Qubit dsDNA HS Assay Kit (Life Technologies); quality checked using an Agilent DNA 1000 Kit (Agilent Technologies); then normalised and pooled. Paired end sequencing (2 x 300 bp reads) of the pooled libraries was conducted on the Illumina MiSeq benchtop sequencing platform (Illumina) and individual indexed reads were demultiplexed using the Local Run Manager Generate FASTQ Analysis Module (Illumina). Library preparation and sequencing was performed by the University of Southampton Environmental Sequencing Facility, and all kits were used in accordance with manufacturers instructions
ATHCWT1