Multi-nucleic acid co-labeling support, preparation method therefor, and application thereof

Abstract

Provided are a variety of nucleic acid co-labeling supports, a preparation method therefor, and an application thereof. The supports comprise a support body, and a variety of nucleic acid labels located on the surface and / or inside of the support body. The nucleic acid labeled on a single support at least comprises: one or more first nucleic acid labels having a function that at least comprises of trapping a specific compound in a reaction system onto the surface of the support; and one or more second nucleic acid labels having a function that at least comprises participating in a specified biochemical reaction process of the specific compound trapped onto the surface of the support. The foregoing variety of nucleic acid co-labeled supports can be used for 5'-terminus single-cell RNA expression profile analysis, construction of a 5'-terminus single-cell VDJ library for a microwell array platform, construction of a 3'-terminus single-cell RNA library, construction of a single-cell transcriptome library, single-cell multi-omics research, multiplex PCR and / or construction of a multiplex PCR sequencing library, etc.

Description

technical field [0001] The invention relates to a multi-nucleic acid co-labeling support and its production method and application. Background technique [0002] Traditional nucleic acid molecular reactions, including nucleic acid hybridization, extension, amplification and other reactions, are all carried out in the liquid phase. The liquid phase provides a uniform and stable environment for the nucleic acid and enzyme reactions involved in the reaction to maximize the output. With the deepening of biological research, scientists have discovered that linking the nucleic acid or enzyme involved in the reaction to the solid-phase surface can give the nucleic acid spatial position information to facilitate purification, separation, detection and analysis. Therefore, more and more solid-phase nucleic acids have been developed. The reaction is used for nucleic acid sequence analysis and nucleic acid quantification, such as nucleic acid chip technology where oligonucleotides are ...

AI Technical Summary

Problems solved by technology

[0007] The droplet method completely isolates cells and label microbeads from other cells and microbeads through water-in-oil, effectively reducing the possibility of cross-contamination; at the same time, in addition to realizing the 3' RNA expression profile library, the droplet scheme can also separate cells Tags and molecular tags are coupled with template-switching sequences to achieve 5' single-cell RNA expression profile sequencing; however, due to the instability and suspension characteristics of the droplets themselves, the single-cell library construction scheme based on the droplet method is difficult to obtain when the RNA is captured by cells. The medium cannot be changed before and after labeling, which reduces the possibility of further complex reactions, especially the lack of position information of the label beads

The microplate method avoids the problem of probability collisions affecting the capture efficiency in 10X, and has better cell capture efficiency; the labeled microbeads have a fixed position after falling into the microwell, and more liquid replacement operations can be performed; however, due to The microwell is a semi-closed structure with an open top that will cause cellular RNA to diffuse out of the hole, so all current microwell methods can only construct 3' single-cell RNA expression profile libraries

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