RNA-seq  Analysis  Reveals the  Response of Tribolium castaneum to the Eugenol Exposure
TotalRNA was processed by the method of mRNA enrichment or rRNA removal to enrich the Mrna;The obtained RNA was segmented by interrupting buffer, and the random N6 primers were reverse-transcribed, and then the two strands of cDNA were synthesized to form double-stranded DNA;The end of the synthesized double-stranded DNA is smoothed and phosphorylated at the 5 'end. The 3' end forms A sticky end with an "A" protruding, and then A bubble-like connector with A "T" protruding at the 6' end is connected;The PCR product was thermally denatured into a single strand, and then a bridging primer was used to loop the single stranded DNA to obtain a single stranded circular DNA library.