Array and method for detecting spatial information of nucleic acids

A method for generating a nucleic acid array for detecting spatial information of a nucleic acid in a biological sample, which comprises the following steps:

• (1) providing multiple kinds of carrier sequences, each kind of carrier sequence comprises a plurality of copies of the carrier sequence, and the carrier sequence in the direction from 5′ to 3′ comprises a positioning sequence and a first immobilization sequence,
• the positioning sequence has a unique nucleotide sequence corresponding to the position of the kind of carrier sequence on the array;
• the first immobilization sequence allows annealing to its complementary nucleotide sequence and initiating an extension reaction;
• (2) ligating the multiple kinds of carrier sequences to a surface of a solid support;
• (3) providing a first primer, and perform a primer extension reaction by using the carrier sequence as a template, so that a region of the first immobilization sequence and the positioning sequence of the carrier sequence forms a double strand, wherein the strand that hybridizes to the carrier sequence is a first nucleic acid molecule, the first nucleic acid molecule in the direction from 5′ to 3′ comprises a complementary sequence of the first immobilization sequence and a complementary sequence of the positioning sequence;
• wherein, the first primer comprises a first immobilization sequence complementary region at its 3′ end, the first immobilization sequence complementary region comprises a complementary sequence of the first immobilization sequence or a fragment thereof, and has a free 3′ end;
• and wherein in step (1), the multiple kinds of carrier sequences are provided by the following steps:
• (i) providing multiple kinds of carrier sequence templates, the carrier sequence template comprising a complementary sequence of the carrier sequence;
• (ii) perform a nucleic acid amplification reaction by using each kind of carrier sequence template as a template, to obtain an amplification product of each kind of carrier sequence template, the amplification product comprising a plurality of copies of the carrier sequence;
• and wherein said each kind of carrier sequence comprising the same positioning sequence occupies an area having a diameter of less than 1 micrometer.