< Return to "NEBNext Ultra II Directional RNA Library Prep: Maximizing Transcript Coverage"
 
 
 

Uniformity of Coverage across ERCC-00002 and ERCC-00145
Dir_RNA_Transcript_Figure8
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). Coverage across transcripts ERCC-00002 (A) and ERCC-00145 (B) were assessed by mapping reads directly to the ERCC sequences and assessing coverage using bedtools cov in 100 bins along the transcript length. Libraries prepared using the NEBNext Ultra II Directional RNA kit provided good coverage across the transcripts at all input amounts.

One or more of these products are covered by patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information, please email us at [email protected]. The use of these products may require you to obtain additional third party intellectual property rights for certain applications.

ILLUMINA®, TRUSEQ® and NEXTSEQ® are registered trademarks of Illumina, Inc.
SPRISELECT® is a registered trademark of Beckman Coulter, Inc.
RIBO-ZERO™ is a trademark of Illumina, Inc.
KAPA™ is a trademark of Kapa Biosystems

Available Kits:

NEBNext_portal_product_icon 

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.

NEBNext_portal_product_icon 

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.