The invention discloses a construction method of a sequencing
library for
pathogenic microorganism detection. The method comprises the following steps: S1, extracting
DNA and
RNA from a sample, and carrying out reverse transcription on tbe
RNA; S2, adding a double-strand synthesis reaction solution and a double-strand synthetase into step S1 to make
RNA / cDNA composite double strands form double-strand cDNA; S3, purifying; S4, adding
a DNA interruption repair reaction solution and
a DNA interruption
repair enzyme mixture into the purified product obtained in step S3, and performing fragmentation, terminal repair and A
tail addition on the product at the same time; S5, directly adding a
linker, a
linker reaction liquid and ligase into the fragmented solution without purifying, and performinglinker connection to obtain a
linker connection product; S6, purifying; S7, carrying out
library amplification; and S8, purifying the amplification product in step S7 to obtain the original
DNA / RNA sequencable
library. The method is short in time; and when the RNA is subjected to one-strand synthesis and two-strand synthesis, only RNA is acted,
DNA existing in a
system is not interfered, and after RNA is converted into double-strand cDNA, RNA and double-strand DNA are fragmented together, so that the effect of co-building the library in one process is truly achieved.