Stem-loop adapters, polynucleotide libraries for guiding ribonucleic acid production, and their application methods

Abstract

(Topic) An adapter for binding polynucleotide fragments to a genome editing system, used to manufacture a comprehensive guide RNA collection for the target genome, and providing the guide RNA collection obtained using these collections and their applications. (Solution) An adapter having the structure described in A). A) Having the structures described in (i) to (iii): (i) A polynucleotide chain having the sequences described in (i)-1 to (i)-3: (i)-1: Pre-primer corresponding sequence (i)-2: Restriction enzyme recognition site sequence for cutting the spacer region of the guide RNA (gRNA) from the 5' side of the polynucleotide fragment bound to the adapter (i)-3: Restriction enzyme recognition site sequence for cutting the adapter having the structure described in A) described above from the polynucleotide fragment (ii) A polynucleotide chain having a sequence complementary to the sequence described in (i) described above (iii) A double-stranded single-stranded polynucleotide chain for loop formation connecting the 5' end of the aforementioned (i) and the 3' end of the aforementioned (ii) described above.