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NEBNext Ultra II Directional RNA libraries provide uniform GC content distribution, at a broad range of input amounts
Dir_RNA_Libraries_Figure4
Ribosomal RNA (rRNA) was depleted from Human Universal Reference RNA (Agilent #740000) with recommended amounts of ERCC RNA Spike-In Mix I (Thermo Fisher Scientific #4456740) and libraries were made using 1 µg, 100 ng and 5 ng input with NEBNext Ultra II Directional RNA Kit (plus the NEBNext rRNA Depletion Kit (Human/Mouse/Rat)), and using 1 µg and 100 ng input with Illumina TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero™ Gold, Kapa Stranded RNA-Seq Kit with RiboErase and Kapa HyperPrep Kit with RiboErase. Libraries were sequenced on an Illumina NextSeq® 500 using paired-end mode (2x76 bp). Reads were mapped to the hg19 reference genome. GC content distribution for each library was calculated using mapped reads. Ultra II Directional RNA libraries had uniform GC content distribution across a range of input amounts.

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Available Kits:

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NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina®
Includes optimized mixes for 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 Directional RNA Library Prep with Sample Purification Beads
Includes optimized mixes for 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.