Ammonolysis method and ammonolysis composition

Abstract

The application of the invention provides a method for performing ammonolysis after DNA solid phase synthesis. The method comprises the following steps of (1) adding an ammonolysis composition to a synthesizing column including a solid phase carrier for synthesizing DNA fragments; (2) placing the synthesizing column in a sealable container loaded with ammonolysis buffer solutions, and maintainingthe synthesizing column to be above the liquid surface of the ammonolysis buffer solutions; (3) performing microwave treatment on the sealable container so that the solid phase carrier is in the steamatmosphere of the ammonolysis buffer solution; and (4) taking out the synthesizing column, treating the solid phase carrier with an eluent, and collecting effluent. The invention further provides anammonolysis composition for ammonolysis. According to the ammonolysis method provided by the invention, only a small quantity of ammonolysis reagents are used, cross infection can be avoided, reactiontime can be shortened, cleaning with cleaning solvents is convenient, high-voltage equipment is not needed, and industrialization and automation use are facilitated.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to an ammonolysis method related to DNA solid-phase synthesis. The invention also relates to an ammonolysis composition for use in ammonolysis. Background technique [0002] DNA synthesis (especially DNA primer synthesis) is mostly based on solid-phase synthesis technology, through the cycle of four steps of deprotection, condensation, oxidation and capping, the phosphoramidite monomers are connected one by one in the order from 3' end to 5' end onto the surface of a controlled pore glass sphere (CPG). The 5' terminal hydroxyl of each phosphoramidite monomer is protected by a dimethoxytriphenyl (DMT) group, and the active amino groups on the base are respectively composed of acetyl (Ac), benzoyl (Bz), di Nitrogen methyl formamido group (dmf) and other groups to protect. Among them, the DMT group is acid-labile and can be removed in a solution such as 3% trichloroacetic acid in dichl...

AI Technical Summary

Problems solved by technology

This method has the following disadvantages: 1) the ammonolysis reagent is easy to volatilize in the process and causes poor repeatability; 2) repeated cooling, the operation is too complicated, and the overall time-consuming is long; 3) the primers cannot be cleaned after the reaction ends, and It takes a long time to drain the mixture of water and methanol; IV) When using a 96-well plate or a 384-well plate for large-scale primer synthesis, the compatibility is poor and cross-contamination is prone to occur

Although the post-treatment link of this method is relatively simple, the ammonolysis method has a high demand for equipment

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