WG0R-220126-34
The second batch of samples were sequenced by Element Biosciences. The nucleic acid samples underwent purification identical to that of HudsonAlpha Discovery, using the RNA Clean and Concentrator Magbead Kit with DNase digestion (Zymo Research, R1082), which cuts both double-stranded and single-stranded DNA. The resulting elution of RNA was quantified using a Qubit 4.0 Fluorometer (ThermoFisher, Q33238), using 10ng of total input for the library preparation. The sequencing libraries were generated using the NEBNext rRNA Depletion Kit (Human/Mouse/Rat) and NEBNext Ultra II DNA Library Prep Kit (New England Biolabs, E6310X, E7645L) for Illumina, following the vendor protocol with two modifications. First, Element Biosciences adapters and unique dual indexes (Element Biosciences, 830-00005) were used instead of NEB recommended Illumina adapters and indexes. Second, because of this change, the USER treatment step was omitted. Completed libraries were quantified by Qubit or equivalent and run on a Bioanalyzer or equivalent for size determination. Libraries were pooled and then circularized and quantified using the Element Elevate Library Circularization Kit (Element Biosciences, 830-00001). Sequencing was performed using the Element AVITI sequencing instrument (Element Biosciences, AVITI), using 2 x 75 bp high output kits (Element Biosciences, 860-00004) run in HD Expert Mode.