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Uniformity of Coverage across the DAM1 transcript
Dir_RNA_Transcript_Figure4
Poly(A)-containing mRNA was isolated from Human Universal Reference RNA
(Agilent #740000), and libraries were made using the NEBNext Ultra II Directional RNA Kit (plus the NEBNext Poly(A) mRNA Magnetic Isolation Module), Kapa Stranded mRNA-Seq Kit, Kapa mRNA HyperPrep Kit and Illumina TruSeq Stranded mRNA Kit. Libraries were sequenced on an Illumina NextSeq® 500 using paired-end mode (2x76 bp). Coverage across the length of this individual transcript (ENST00000369541.3; DAM1) was assessed by mapping reads directly to the GENCODE v25 transcripts and examining 100 bp bins along the transcript length. NEBNext Ultra II Directional RNA libraries provided coverage across the entire length of the transcript, even as input was decreased from 1 µg to 10 ng.

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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.