Nucleic Acid Synthesis Methods

Abstract

The invention provides a nucleic acid synthesis method. The synthesis method comprises: splitting the nucleic acid to be synthesized into multiple groups of fragments according to sequence order and synthesizing respectively to obtain multiple groups of nucleic acid fragments, each group of nucleic acid fragments containing 2 to 5 bases; The fragments are joined in sequence order to obtain nucleic acids. This application creatively proposes a new synthesis strategy of multi-base short fragment coupling instead of single-base coupling. Multiple short fragments containing 2-5 bases are first synthesized, and then sequentially spliced ​​using solid-phase synthesis. . Multiple fragments can be synthesized in parallel at the same time, and it is not limited by the loading capacity of the solid phase carrier, and a large number of high-purity short fragments can be easily obtained. Splicing in solid phase reduces the original connection steps by at least 1 / 2, greatly shortens the reaction time, improves the reaction efficiency and overall yield, and avoids the generation of N-1 non-target fragments in traditional synthesis. The purity of the product is improved.

Description

technical field [0001] The present invention relates to the field of nucleic acid synthesis, in particular to a nucleic acid synthesis method. Background technique [0002] At present, solid-phase synthesis of nucleic acids is performed using trivalent phosphorus chemistry, through four-step cyclic reactions, and one base is ligated at a time. The advantage of this method is that it is fast and efficient. But at the same time, fragments with incomplete coupling exist, that is, there is an N-1 problem. Bring great trouble to follow-up purification. Taking Spinraza as an example, the traditional synthesis method of Spinraza also uses a solid phase carrier to connect one base at a time, which requires 18 steps of ligation reactions to achieve. Therefore, not only the synthesis steps are longer, but also short fragments such as N-1 exist in the synthesis process, making purification difficult. [0003] Therefore, there is still a need to provide a relatively fast and efficie...

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Problems solved by technology

[0004] The main purpose of the present invention is to provide a nucleic acid synthesis method to solve the problems of long synthesis steps and low efficiency in existing methods

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