Method for simply and quickly preparing ready-to-use esiRNA

Abstract

The invention relates to a preparation method of a small RNA (Ribose Nucleic Acid), in particular to a new method for simply and quickly preparing ready-to-use esiRNA at a low cost. The method has the beneficial effects that a chip integrating a macromolecule microsphere (macromolecule block) is adopted to simply and quickly prepare the esiRNA at a low cost and is utilized for amplification, transcription and enzymic digestion on a polymer bracket, so that liquid transfer, purification and other operation steps are simplified; the preparation method of the esiRNA is reduced from the existing 6 steps to 3 steps provided by the invention, so that the preparation process of the esiRNA is greatly simplified; and meanwhile, the chip is utilized to cut and normalize the quantity of the esiRNA product, so that the esiRNA can be directly used for the research of loss-of-function caused by gene silencing without further treatment.

Description

technical field [0001] The present invention relates to a preparation method of small RNA, in particular to a simple, fast and low-cost new method for preparing ready-to-use esiRNA. Background technique [0002] RNA interference (RNA interfering, RNAi) is a phenomenon in which double-stranded RNA (double-stranded RNA, dsRNA) induces efficient and specific degradation of homologous mRNA. It is an ancient and evolutionarily highly conserved gene expression regulation mechanism in the biological world. In 1998, researchers discovered that injecting dsRNA into C.elegans or Drosophila can specifically inhibit gene expression, and called this phenomenon of inhibiting specific gene expression triggered by dsRNA as RNAi. Subsequently, the basic research and application of RNAi quickly became one of the hot areas of life science. RNAi research has made a breakthrough, and was rated as one of the top ten scientific advances in 2001 by the "Science" magazine, and ranked first in the t...

AI Technical Summary

Problems solved by technology

[0004] However, the preparation of esiRNA in the prior art is a rather complicated process, including at least 6 steps: PCR amplification, transcription, enzyme digestion, purification, esiRNA quantification and normalization ( figure 1 ), these steps include the need to transfer, purify, quantify, and normalize liquid samples, requiring the use of expensive instruments such as liquid handling systems, centrifuges, and spectrometers

These tedious and expensive work have seriously affected the large-scale preparation and application of esiRNA

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