5' DNA Adenylation Kit

5' DNA Adenylation Kit is specifically designed for the adenylation modification of 5'-phosphorylated single-stranded DNA (ssDNA) at the 5' end. This kit contains Mth RNA Ligase (an RNA ligase derived from thermophilic archaea) and ATP. During the adenylation reaction, 5'-phosphorylated single-stranded DNA (5'-pDNA) serves as the substrate. The Mth RNA Ligase, which possesses adenylation activity, hydrolyzes ATP into AMP and PPi, and subsequently transfers the AMP to the 5'-phosphate group of the ssDNA, ultimately generating 5'-adenylated ssDNA (5'-AppDNA).

This kit offers a simple and efficient one-step reaction for quantitative adenylation, typically converting 95% of pDNA to AppDNA, eliminating the need for product purification. It is more convenient than current chemical or enzymatic methods. The reaction temperature of 65°C effectively reduces secondary structure formation, and is suitable for reaction systems ranging from pmol to µmol substrate amounts. This product is commonly used to generate adenylated linkers for 3'-end ligation to ssRNA/ssDNA with a 3'-hydroxyl group, facilitating applications such as cloning, high-throughput sequencing library preparation, or PCR detection.