This kit is an RNA library construction kit based on a novel transposable enzyme system, which is suitable for Illumina sequencing platform. After the synthesis of A strand of cDNA, a novel transposase is used to fragment and add splicing to the RNA/DNA heterozygous strand, which replaces the steps of synthesis, fragmentation, end repair plus A and splicing to the two-strand cDNA in the process of conventional RNA library construction. The library can be completed after PCR amplification and purification. Greatly simplify the operation process and shorten the library construction time, with a single library construction time of no more than 120 minutes. Moreover, in the sample input range of 1-50 ng, without considering the input amount of template, the optimized kit system can meet the needs of using only the same set of reagents and the same process to complete the library construction, ensuring the efficiency and stability of library construction.