Labeling and analysis method for single-cell nucleic acid

A method for generating a population of labeled nucleic acid molecules, which comprises the following steps:

• (1) providing:
• a sample containing one or more cells, and a nucleic acid array;
• wherein, the sample is a single-cell suspension;
• the cell comprises (e.g., on its surface) a first binding molecule;
• the nucleic acid array comprises a solid support, the solid support comprises (e.g., on its surface) a first label molecule, and the first binding molecule is capable of forming an interaction pair with the first label molecule;
• and, the solid support further comprises a plurality of microdots, the size (e.g., equivalent diameter) of the microdots is less than 5 μm, and the center-to-center distance between adjacent microdots is less than 10 μm;
• each microdot is coupled with one kind of oligonucleotide probe, each kind of oligonucleotide probe comprises at least one copy;
• the oligonucleotide probe in the direction from 5′ to 3′ comprises or consists of:
• a consensus sequence X1, a tag sequence Y and a consensus sequence X2, wherein,
• oligonucleotide probes coupled to different microdots have different tag sequences Y;
• (2) contacting the one or more cells with the solid support of the nucleic acid array, whereby each cell individually occupies at least one microdot in the nucleic acid array (i.e., each cell is individually contacted with at least one microdot in the nucleic acid array), and allowing the first binding molecule of the cell to interact with the first label molecule of the solid support;
• wherein,
• before or after contacting the one or more cells with the nucleic acid array, performing a pretreatment, including reverse transcription, on an RNA (e.g., an mRNA) of the one or more cells to generate a first nucleic acid molecule population;
• and,
• (3) associating the first nucleic acid molecule population derived from each cell obtained in the previous step with an oligonucleotide probe coupled to the microdot occupied by the cell from which the first nucleic acid molecule population is derived, thereby generating a second nucleic acid molecule population labeled with the tag sequence Y.