Description

5'-rApp is a nucleic acid molecule that contains a 5'-5'-adenosine diphosphate cap. It possesses a high-energy pyrophosphate bond and serves as an activated form of nucleic acid. Through 5'-adenylation modification, short oligonucleotides can be linked to the 3'OH end of different sequences without relying on ATP. This modification is commonly used as a 3' adapter during the construction of miRNA or single-stranded DNA libraries with a hydroxyl group at the 3' end, reducing self-ligation between molecules. With the widespread application of next-generation sequencing technologies in high-throughput sequencing, RNA high-throughput sequencing techniques have also become increasingly mature. In the construction of small RNA libraries, 5'-adenylation modification (represented by 5'-rApp, where "pp" denotes a diphosphate linkage from 5' to 5' end) of adapters is necessary to prevent self-ligation of RNA molecules and minimize the formation of RNA chimeras in the library. 5'-adenylated oligos can connect 3'-OH ends of different sequences and are commonly used for linking 3' hydroxyl-ended single-stranded DNA or miRNA, frequently employed in library construction.

Currently, there are two main methods for synthesizing 5'-rAppDNA: enzymatic synthesis and chemical synthesis. Enzymatic synthesis involves complex steps, including the enzymatic breakdown of ATP into AMP and PPi, followed by the transfer of AMP to the 5' phosphate group of the oligonucleotide, resulting in adenylation. On the other hand, chemical synthesis directly connects the rApp modification to the 5' end of the oligonucleotide without the involvement of enzymes or ATP. Chemical synthesis offers advantages such as high purity and low cost. We provide 5' adenylation modification services, offering chemically synthesized 5'-adenylated oligonucleotides (Oligos), suitable for applications such as miRNA library construction and related scenarios.

Advantage

Our rApp adenylated nucleic acids undergo meticulous mass spectrometry analysis and HPLC purification, ensuring exceptional purity and high accuracy of the synthesized products. This means that you can obtain high-quality products that provide a reliable foundation for your research and applications.

Other Applications 

• Activated Substrates for Exonuclease/Endonuclease in vitro Selection: 5'-App oligonucleotides with site-specific modifications are used for the development of in vitro selection of exonucleases/endonucleases and the construction of specific long RNAs for RNA structure-function studies.
• Activation of 5'-End Pyrimidine-Rich RNA: By introducing 5'-adenylation modification, the 5'-end pyrimidine-rich RNA can be activated for efficient ligation, overcoming the challenge of synthesizing such RNA through in vitro transcription.
• 5'-End Labeling: By oxidizing 5'-App nucleotides to aldehydes, followed by reductive amination, fluorescent dyes or other biophysical probes can be introduced at the 5'-end. This labeling strategy can be used for situations where specific 5'-end modifications are not feasible or desired using solid-phase synthesis.

These applications demonstrate the versatility and flexibility of rApp in nucleic acid research, providing effective tools to achieve specific experimental goals.