The RNA-seq experiment was performed as described previously (20 (link)). Paramagnetic beads coupled with oligo d(T) are combined with total RNA to isolate poly(A)+ transcripts based on NEBNext Poly(A) mRNA Magnetic Isolation Module manual. All remaining steps for library construction were used according to the NEBNext Ultra II RNA Nondirectional Library Prep Kit from Illumina. Illumina 8-nt dual-indices. Samples were pooled and sequenced on a HiSeq with a read length configuration of 150 PE. Details and references are provided in the Supplementary Materials and Methods.
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Poly(A)+ RNA Isolation and Nondirectional Library Prep
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Variable analysis
independent variables
- Protocol for isolation of poly(A)+ transcripts using paramagnetic beads coupled with oligo d(T)
- Sequencing of the constructed libraries on HiSeq (read length configuration of 150 PE)
- Total RNA as the starting material
- Use of the NEBNext Poly(A) mRNA Magnetic Isolation Module and NEBNext Ultra II RNA Nondirectional Library Prep Kit from Illumina for library construction
- No positive or negative controls are explicitly mentioned in the provided information.
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