NEXTflex™ Small RNA Sequencing Kit

Library prep for sequencing on Illumina platforms

Catalog# Product Name Quantity US List Price
5132-01 NEXTflex™ Small RNA-Seq Kit
24 rxns $1,350
5132-02 NEXTflex™ Small RNA-Seq Kit 48 rxns $2,250
513301 NEXTflex™ Small RNA Barcodes Primers - 12 (Set A)
96 rxns $400
513302 NEXTflex™ Small RNA Barcodes Primers - 12 (Set B)
96 rxns $400
513303 NEXTflex™ Small RNA Barcodes Primers - 12 (Set C)
96 rxns $400
513304 NEXTflex™ Small RNA Barcodes Primers - 12 (Set D) 96 rxns $400

    • Up to 48 barcodes available for multiplexing
    • Utilizes AIR™ Ligase, a highly efficient truncated T4 RNA Ligase, for greater sequencing depth
    • Simplified workflow reduces hands-on time
    • Flexible barcode options
    • Barcoded PCR Primers for multiplexing contain embedded index sequence
    • Automation-friendly workflow is compatible with liquid handlers
    • Functionally validated with the Illumina sequencing platforms  




    The patent pending NEXTflex™ Small RNA Sequencing Kits provide an easy, flexible, cost-effective solution for generating single and multiplexed libraries from small RNA using Illumina sequencing platforms. This kit contains a specialized 3’ adenylated adapter that specifically ligates onto microRNAs and other small RNAs containing a 3’ hydroxyl group. After adapter ligation and reverse transcription, libraries are amplified using a barcoded primer, allowing up to 48 samples to be multiplexed and sequence variations interrogated. The NEXTflex™ Small RNA Sequencing Kit incorporates AIR™ Ligase, an enhanced, truncated T4 RNA Ligase, which increases the efficiency with which small RNAs are tagged with adapter, giving greater sequence depth. The innovative NEXTflex adapter ligation technology that is incorporated into the NEXTflex Small RNA Sequencing Kit greatly reduces the adapter dimer band giving better quality and a higher number of sequence reads with less contamination from adapter dimer. Using a specialized RT primer and buffer strategy, 3’ and 5’ adapter dimers will no longer impede your research by taking up valuable sequencing reads.


    The NEXTflex™ Small RNA Sequencing Kit on its own contains all the material to prepare 24 or 48 singleplex libraries. In combination with the NEXTflex™ Small RNA Barcode Primer Kits, 48 samples can be profiled and studied simultaneously. The NEXTflex Small RNA Barcode Primer Kits contain 8 reactions each of 12 unique barcodes for mulltiplexing. In addition to small RNA sequencing, the kit is compatible with RIP-Seq and CLIP-Seq protocols.


    For larger volume requirements, customization and bulk packaging is available. For increased flexibility individual reagents (non-master mixed) are also available. Please contact for further information. 


    Reducing Bias in Small RNA-Seq Library Prep:

    Read our blog post, Reducing Bias in Small RNA-Seq Library Prep,  describing new technology available to reduce bias introduced by ligation in small RNA sequencing library preparation.

    Selected Citations:

    Ahmed R, Chang Z, Younis A.E. et al. (2013) Conserved miRNAs Are Candidate Post-Transcriptional Regulators of Developmental Arrest in Free-Living and Parasitic Nematodes. Genome Biol Evol (2013) 5 (7): 1246-1260. doi: 10.1093/gbe/evt086

    Jayaprakash, A. D., Jabado O., Brown, B. D. and Sachidanandam, R. (Sept 2, 2011), Identification and remediation of biases in the activity of RNA ligases in small-RNA deep sequencing Nuc Acid Res, 1–12. doi:10.1093/nar/gkr693

    Langlois, R. A. et al. (Nov 15, 2011), In Vivo Delivery of Cytoplasmic RNA Virus-derived miRNAs. Molecular Therapy. doi:10.1038/mt.2011.244

    Maczuga, P. et al. (Jan 2013) Embedding siRNA sequences targeting Apolipoprotein B100 in shRNA and miRNA scaffolds results in differential processing and in vivo efficacy. Molecular Therapy (2013); 21 1, 217-227. doi: 10.1038/mt.2012.160

    Nakashe, P. et al. (2011) Adapter Dimer Reduction in High-Throughput microRNA Profiling. OMICS 1(1): 6-11.

    Sorefan, K. et al. (2012) Reducing sequencing bias of small RNAs. Silence. doi:10.1186/1758-907X-3-4

    Velthut-Meikas A, Simm J, Tuuri T, Tapanainen JS, Metsis M, Salumets A (2013) Small RNA-seq of Human Granulosa Cells Reveals miRNAs in FSHR and Aromatase Genes. Mol. Endocrinol. 2013; 27:1128-1141.


    Smith, O. et. al. (2013) A complete ancient RNA genome: identification, reconstruction and evolutionary history of archaeological Barley Stripe Mosaic Virus. Scientific Reports. doi:10.1038/srep04003 


    Kit Specs

    The NEXTflex™ Small RNA-Seq Kits contain the reagents to prepare 24 or 48 small RNA samples for Illumina®-compatible next-generation sequencing. The shelf life of all reagents is 12 months when stored properly at -20°C. Ships on dry ice.