The invention discloses a construction method of a
single cell transcriptome sequencing
library and application of the construction method. The construction method includes the following steps of preparing a cDNA sample from a single
cell and conducting fragmentized
library construction on the cDNA sample to obtain the
transcriptome sequencing
library of the single
cell, wherein fragmentation is conducted on the cDNA sample with a physical breaking method in the step of constructing the fragmentized library, and the step for purifying the fragmentized cDNA sample is not needed. According to the method, fragmentation is conducted on the sample of the single
cell with the physical breaking method, and therefore fragments obtained through breaking are small and relatively uniform, and the peak width of the constructed
single cell transcriptome sequencing library is concentrated; the purification step is not needed for the fragmentized sample, sample loss is reduced, construction of the
single cell transcriptome sequencing library of the micro-sample can be achieved, stability of output of data volume can be improved, and therefore the reliability of large-scale and high-
throughput sequencing of the single cell
transcriptome sequencing library is improved.