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Uniformity of Coverage across ERCC-00004 and ERCC-00074
Nondir_RNA_Transcript_Figure3
Ribosomal RNA was depleted from Human Universal Reference RNA (Agilent #740000) with recommended amounts of ERCC RNA Spike-In Mix I (Thermo Fisher Scientific #4456740) using the NEBNext rRNA Depletion Kit (Human/Mouse/Rat). Libraries were then prepared using the NEBNext Ultra II RNA Kit. Libraries were sequenced on an Illumina NextSeq® 500 using paired-end mode (2x76 bp). Coverage across transcript ERCC-00004 (A) and ERCC-00074 (B) were assessed by mapping reads directly to the ERCC sequences and assessing coverage using bedtools in 100 bins along the transcript length. Libraries prepared using the NEBNext Ultra II RNA Kit provided good coverage across the transcripts at all input amounts.

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Available Kits:

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NEBNext® Ultra™ II RNA Library Prep Kit for Illumina®
Includes optimized mixes for non-directional RNA library preparation (fragmentation, cDNA synthesis, end repair/dA-tailing, adaptor ligation and PCR enrichment steps) for sequencing on the Illumina platform.

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NEBNext® Ultra™ II RNA Library Prep with Sample Purification Beads
Includes optimized mixes for non-directional RNA library preparation (fragmentation, cDNA synthesis, end repair/dA-tailing, adaptor ligation and PCR enrichment steps) plus SPRIselect® beads for size selection and cleanup.