Adenylator™ DNA/RNA 5’-Adenylation Kit
- Produce 3 nmol of 5’-adenylated DNA or RNA oligos, triple the output of competitor kits.
- Avoid off-target effects like concatemerization and circularization seen in ligase-based adenylation methods.
- Adenylate oligos in a one-step, one-hour reaction instead of waiting weeks for purchased adenylated oligos.
- Scale up reactions as desired with no impact on adenylation efficiency.
Product is for research use only (RUO)
Product Description:
The Adenylator™ DNA/RNA 5’-Adenylation Kit enables ligation-free, efficient, 5’-end adenylation of 5’-monophosphorylated DNA and RNA oligonucleotides. The kit provides a powerful, catalytic and low-temperature alternative to 5’-adenylation methods that use traditional thermostable RNA ligase, providing triple the output of competitor kits. Unlike Mth RNA Ligase, which relies on high temperature (65°C) to minimize oligo circularization and concatemerization events, Adenylator™ Enzyme performs robustly at 37°C and cannot catalyze unwanted oligo self-ligation.
Each 10-reaction kit adenylates 3 nmol of DNA or RNA oligos in a one-step, one-hour, scalable workflow. The resulting 5’-adenylated oligos are compatible with T4 RNA Ligase 2 truncated or truncated/K227Q-based reactions for ligation to the 3’-end of RNA. Adenylator™ adenylated DNA oligos can be used in next-generation sequencing workflows (e.g., small RNA library prep and Native elongating transcript sequencing [NET-seq]) and adenylated RNA oligos can be used as an input into RNA circularization workflows or in a variety of RNA studies.
Product Performance:
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 visualized 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 catalyze unwanted oligo self-ligation. A 23-nt phosphorylated DNA oligo was adenylated using Adenylator™ Enzyme and Mth RNA Ligase and visualized 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 circularized oligo (blue arrow) ligation byproducts. Adenylation using Adenylator™ Enzyme (T-A) cannot form substrate concatemers or circularize oligo substrates, resulting in adenylated oligos without byproducts.
