Single molecule sequencing method, device, system, and application

The present invention provides a single molecule sequencing (SMTS) method, device, system, and application, and the method comprising: (i) combining a template nucleic acid having a first optical detection marker at an end with a primer to obtain a first complex; (ii) imaging the first complex to obtain a first image; (iii) mixing the first complex, a polymerase, and at least one nucleotide having an optical detection marker, and obtaining an extended product by a polymerase reaction; (iv) imaging the extended first complex to obtain a second image; (v) removing a breakable group from the nucleotide having the optical detection marker of the extended product to obtain a second complex; and (vi) repeating step (ii) to (v) one or more times to identify a sequence of the template nucleic acid. The single molecule sequencing method provided by the present invention achieves single molecule sequencing via a primer capturing a template nucleic acid.