Abstract: Provided are a method for constructing a library based on an RNA sample and uses thereof. The method includes: step 1 of subjecting the RNA sample to a reverse transcription reaction to obtain DNA-RNA hybrid strands; step 2 of performing reaction of the DNA-RNA hybrid strands with an endoribonuclease, a first DNA polymerase, a second DNA polymerase, and dATPs to obtain a double-stranded DNA added with dA-tail, where the first DNA polymerase has a 5?-3? exonuclease activity and a 3?-5? exonuclease activity, and the second DNA polymerase has no 3?-5? exonuclease activity; step 3 of ligating the double-stranded DNA added with dA-tail and a sequencing adaptor to obtain a ligated product; and step 4 of subjecting the ligated product to PCR amplification to obtain a sequencing library.

Abstract: Provided are an RNA library construction method and a specialized kit thereof. The method for preparing an RNA library includes the following steps: extracting RNA and performing fragmentation; adding a tail to 3? end; ligating an adaptor to 5? end and hybridizing with a DNA probe mixture; the DNA probe mixture is composed of several DNA probes that are reverse complementary to an RNA that is expected to be removed; removing RNA from the hybrid and removing DNA; performing reverse transcription and PCR amplification to obtain a library solution. The present invention combines polyA tailing and 5?end ligation. The RNA content in the system can be quickly increased by polyA tailing, thereby avoiding subsequent purification losses.