ArticleMethodsX2025
An oligo-swapping method: preparation of mismatch repair-monitoring substrate using a nicking endonuclease.
Article in MethodsX, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Mismatch repair (MMR) contributes to accurate DNA replication by eliminating mismatched bases during DNA synthesis. Its importance is underscored by Lynch syndrome, a common hereditary colorectal cancer syndrome caused by MMR gene mutations. Therefore, quantification of MMR activity in human cells is important for diagnostic and therapeutic purposes. To monitor MMR, we established a novel DNA plasmid, pBluescript II NLS-MC-EGFP-tdTomato (pBET2). After processing using our protocol called the oligo swapping method, the mismatch included in the pBET2 enables us to evaluate the MMR proficiency of living cells by detecting specific fluorescent markers. Since the method simply "swaps" short single-stranded DNA to create a single mismatch on a specific site of the plasmid, it is significantly easier and more user-friendly. Briefly, the nicking endonuclease Nt.
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