In high-throughput sequencing (NGS) and reversible termination sequencing (RTS), "making DNA replication controllably stop" is key to achieving precise sequencing.

The core of achieving this "reversible termination" lies in nucleoside triphosphates (dNTP) with 3′-O modified groups.

Traditional dNTPs undergo continuous extension under polymerase action and lack pause control capability.

When research requires real-time observation of each base incorporation process at the single-molecule level, precise chemical modification of dNTP becomes crucial.

This is why 3′-O-azidomethyl makes its appearance.

3′-O-Azidomethyl is a temporary protecting group that can be removed by light or reducing agents.

When it modifies the 3′ hydroxyl group of dNTP:

This cycle of "termination—deprotection—re-extension" is the core mechanism of reversible termination sequencing.

Simultaneously, the azide group can also participate in click chemistry reactions (such as CuAAC reactions), endowing the nucleotide with post-modification and labeling capabilities.

1. Reversible Termination Sequencing: Controls single-base extension, enables base-by-base detection, and improves sequencing accuracy.

2. Enzyme Screening and Polymerase Reaction Studies: Used to evaluate polymerase recognition and catalytic activity towards 3′ modified substrates.

3. Click Chemistry Labeling and Probe Design: The N₃ group enables highly selective CuAAC reactions, facilitating nucleic acid labeling.

✅ Strong Compatibility: The modifying group is small, causing minimal interference to the polymerase, ensuring efficient incorporation and synthesis.

✅ Scarless Restoration: After cap removal, the DNA strand completely restores its natural structure with no residual modification, supporting ultra-long read lengths.

✅ Specific Signal: The azide group has a unique Raman signal with very low biological background interference.

✅ Precise Start-Stop Control: Controllable 3′ end blocking supports high-precision sequencing cycles.

To meet research needs, Shenzhen GeneSeqTools Bioscience & Technology Co. Ltd. introduces the 3′-O-Azidomethyl-dNTP Series, covering five bases (A, C, G, T, U) to facilitate experimental design and system setup.

Products are available in 25 μl / 100 μl / 250 μl specifications, support custom packaging, and flexibly meet different experimental scales and application needs.