DNA Linker Adenylation for RNA-seq & NGS

Reliable 5’ Adenylation for High-quality RNA Libraries and NGS workflows

At Tebubio, we understand that successful RNA-seq and NGS experiments depend on robust, reproducible enzymatic steps and reagents you can trust. That is why we distribute the Adenylator™ DNA/RNA 5’-Adenylation Kit, a high-performance solution designed to simplify and optimise 5’ linker adenylation for demanding sequencing applications.

The Adenylator™ DNA/RNA 5’-Adenylation Kit enables ligation-free, catalytic adenylation of 5’-monophosphorylated DNA and RNA oligonucleotides in a single-step, one-hour workflow. Operating efficiently at 37 °C, this enzyme eliminates the need for high-temperature reactions and avoids common artefacts associated with traditional RNA ligases.

Unlike Mth RNA Ligase, which requires elevated temperatures (65 °C) to reduce circularisation and concatemer formation, Adenylator™ Enzyme cannot catalyse unwanted self-ligation events, ensuring cleaner products and more reliable downstream results.

Product information

• Catalogue number: ADR250610

• Shipping conditions: Cool pack

• Storage temperature: –20 °C

Each 10-reaction kit adenylates up to 3 nmol of DNA or RNA oligonucleotides, offering scalability for both method development and routine library preparation. The resulting 5’-adenylated oligos are fully compatible with:

• T4 RNA Ligase 2 (truncated or truncated/K227Q) reactions

• Small RNA library preparation

• NET-seq (Native Elongating Transcript Sequencing)

• RNA circularisation workflows and advanced RNA studies

This makes the Adenylator™ kit an ideal choice for laboratories seeking higher yield, reduced background and improved reproducibility in sequencing workflows.

Proven Performance You Can Trust

Figure 1. DNA and RNA oligonucleotides are efficiently adenylated using the Adenylator™ DNA/RNA 5’-Adenylation Kit
Phosphorylated DNA (Fig. 1A) and RNA (Fig. 1B) oligonucleotides were adenylated using Adenylator™ Enzyme and visualised on polyacrylamide gels (20% acrylamide, 8 M urea, 1X TBE, stained with SYBR® Gold). The untreated oligos (lane 2) were efficiently adenylated (lane 3) as evidenced by the band shift.

Figure 2: Adenylator™ Enzyme does not catalyse unwanted oligo self-ligation
A 23-nt phosphorylated DNA oligo was adenylated using Adenylator™ Enzyme and Mth RNA Ligase and visualised on a polyacrylamide gel (20% acrylamide, 8 M urea, 1X TBE, stained with SYBR® Gold). The Mth RNA Ligase adenylated DNA oligo (T-M) contains oligo concatemer (red arrows) and circularised oligo (blue arrow) ligation byproducts. Adenylation using Adenylator™ Enzyme (T-A) cannot form substrate concatemers or circularise oligo substrates, resulting in adenylated oligos without byproducts.

Key Benefits for Your Laboratory

• Higher turnover rate: High specificity for p-DNA and p-RNA 5’ ends

• No ligase activity: No off-target concatemerisation or circularisation

• Single-step reaction: Faster protocols, reduced hands-on time

• Cost-effective: Eliminates the need for chemically synthesised adenylated oligos

Your Trusted Distribution Partner

As a specialised life science distributor, Tebubio provides high-performance, validated reagents backed by scientific expertise. Our team can help you choose the right enzymes for your RNA-seq and NGS workflows, ensuring reliable results and faster discovery.

• References

  Article content created by Tebubio using courtesy materials provided by CellScript.

Need Expert Guidance for Your RNA-seq and NGS Experiments?

At Tebubio, our scientific team can help you optimise your 5’-adenylation of DNA and RNA oligos for RNA-seq, small RNA libraries, or NET-seq. Receive tailored advice to maximise adenylation efficiency, minimise unwanted by-products, and ensure reliable, reproducible results for your downstream applications.