The present application relates to the field of
biotechnology and molecular detection, in particular to a
plant and
pathogenic bacteria interaction
gene expression analysis method based on UMI tag, comprising: obtaining interaction sample total
nucleic acid extract; and constructing a
molecular hybridization system with host-specific chimeric blocking probes containing
locked nucleic acid modification and 3' end dideoxy modification, and reverse transcription primers; through temperature rising and denaturation and temperature reduction and annealing, the blocking probes are combined with host high abundance
nucleic acid to form a hybridization blocking complex, and the primers are combined with
pathogenic bacteria nucleic acid;
template switching oligonucleotides with unique molecular identifier sequences and
reverse transcriptase are added, the host extension is blocked, and UMI is introduced into the
pathogenic bacteria complementary chain, and an interaction
expression library is generated; after sequencing, the pathogenic
bacteria effective reads and the host invalid reads are counted, and the effective interaction
signal signal-to-
noise ratio is output; the present application can preferentially capture pathogenic
bacteria transcripts in the high background of host
RNA.