Enrichment of mRNA was performed using 250 ng total RNA and the NEBNext Poly(A) Magnetic Isolation Module (New England BioLabs, Whitby, ON), and cDNA was subsequently synthesized using the NEBNext RNA First Strand Synthesis and NEBNext Ultra Directional RNA Second Strand Synthesis Modules (New England BioLabs).
More...Enrichment of mRNA was performed using 250 ng total RNA and the NEBNext Poly(A) Magnetic Isolation Module (New England BioLabs, Whitby, ON), and cDNA was subsequently synthesized using the NEBNext RNA First Strand Synthesis and NEBNext Ultra Directional RNA Second Strand Synthesis Modules (New England BioLabs). The remaining steps of library preparation were carried out using the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England BioLabs). Libraries were quantified using the Kapa Illumina GA with Revised Primers-SYBR Fast Universal kit (Kapa Biosystems, Wilmington, MA) and average fragment size was determined using a LabChip GX instrument (PerkinElmer, Waltham, MA). Sequencing was carried out using an Illumina NovaSeq 6000 platform (Illumina Inc., San Diego, CA) with 100-bp paired end reads.
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