title: FYP1
design: Sequencing of lncRNA removes ribosomal RNA from total RNA, thus obtaining all lncRNA, and theoretically some mRNA and circRNA. subsequently, the obtained RNA is randomly interrupted, and libraries are constructed according to strand-specific library construction. Strand-specific library construction: the first strand of cDNA was synthesized by reverse transcription in the same way as the NEB common library construction method, the difference is that when synthesizing the second strand, the dTTP in the dNTPs was replaced by dUTP, and the same cDNA end repair, addition of A-tail, joining of sequencing junction, and length screening were also carried out, and the second strand of the U-containing cDNA was degraded by USER enzyme before PCR was carried out to amplify and obtain the library. amplification and library acquisition. Strand-specific libraries have many advantages, such as obtaining more valid information with the same amount of data; obtaining more accurate gene quantification, localization and annotation information; and providing antisense transcripts and expression levels of a single exon in each isoform.