GSM9355035: 1.0 m1Psuedouridine Rep 3; Homo sapiens; RNA-Seq
Column Purification 300 ng of total RNA for each treatment were submitted for sequencing by Source Biosciences (Cambridge). There, samples were poly(A) selected before cDNA generation with the NEB Ultra II DNA Library Preparation Kit (New England Biolabs), targeting 40,000,000 paired end reads of 150 bp lengths - and resulting in libraries which are forward stranded.  For each treatment, 300 ng of total RNA were submitted to Source Bioscience (Cambridge) for sequencing. Samples underwent poly(A) selection followed by cDNA synthesis using the NEBNext Ultra II DNA Library Preparation Kit (New England Biolabs), with a target depth of approximately 40 million 150 bp paired-end reads per sample. Four experimental conditions, m1Ψ100, 5moU25, unmodified mRNA, and mock, in triplicate. Two independent batches of sequencing were performed, one batch containing mock (N=3) and m1Ψ100 (N=3), and a separate batch comprising mock (N=3), 5moU25 (N=3), and unmodified (N=3) mRNA treatments.