Correcting errors in PCR-derived libraries for rare allele detection by reconstructing parental and daughter strand information

Abstract Molecular barcoding methods enable high-sensitivity detection of circulating tumor DNA that is rarely present in liquid biopsy samples. Many methods involve ligation of molecular barcodes to DNA prior to hybridization capture, enabling recovery of starting molecules. Development of polymera...

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Main Authors: Hyeonseob Lim, Soyeong Jun, Taehoon Kim, Ji Hyun Lee, Duhee Bang
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Communications Biology
Online Access:https://doi.org/10.1038/s42003-025-08537-3
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author Hyeonseob Lim
Soyeong Jun
Taehoon Kim
Ji Hyun Lee
Duhee Bang
author_facet Hyeonseob Lim
Soyeong Jun
Taehoon Kim
Ji Hyun Lee
Duhee Bang
author_sort Hyeonseob Lim
collection DOAJ
description Abstract Molecular barcoding methods enable high-sensitivity detection of circulating tumor DNA that is rarely present in liquid biopsy samples. Many methods involve ligation of molecular barcodes to DNA prior to hybridization capture, enabling recovery of starting molecules. Development of polymerase chain reaction (PCR)-based methods could facilitate more cost- and labor- effective detection; however, tracking molecular identity can be difficult, as new barcodes overwrite old barcodes in each cycle. Here, we present a sensitive genotyping method based on a peer-to-peer network-derived identifier for error reduction in amplicon sequencing (SPIDER-seq) and enable molecular identity tracking with PCR-derived libraries using overwritten barcodes. SPIDER-seq detected mutations at frequences as low as 0.125% after only two consecutive general PCR cycles and systematically analyzed the error pattern in the peer-to-peer network. Our method could facilitate the rapid detection of mutations associated with various cancers.
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issn 2399-3642
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spelling doaj-art-8858435e673b4b7ab4e5425a36a686532025-08-20T03:05:56ZengNature PortfolioCommunications Biology2399-36422025-07-018111210.1038/s42003-025-08537-3Correcting errors in PCR-derived libraries for rare allele detection by reconstructing parental and daughter strand informationHyeonseob Lim0Soyeong Jun1Taehoon Kim2Ji Hyun Lee3Duhee Bang4Department of Chemistry, Yonsei UniversityDepartment of Chemistry, Yonsei UniversityDepartment of Chemistry, Yonsei UniversityDepartment of Clinical Pharmacology and Therapeutics, College of Medicine, Kyung Hee UniversityDepartment of Chemistry, Yonsei UniversityAbstract Molecular barcoding methods enable high-sensitivity detection of circulating tumor DNA that is rarely present in liquid biopsy samples. Many methods involve ligation of molecular barcodes to DNA prior to hybridization capture, enabling recovery of starting molecules. Development of polymerase chain reaction (PCR)-based methods could facilitate more cost- and labor- effective detection; however, tracking molecular identity can be difficult, as new barcodes overwrite old barcodes in each cycle. Here, we present a sensitive genotyping method based on a peer-to-peer network-derived identifier for error reduction in amplicon sequencing (SPIDER-seq) and enable molecular identity tracking with PCR-derived libraries using overwritten barcodes. SPIDER-seq detected mutations at frequences as low as 0.125% after only two consecutive general PCR cycles and systematically analyzed the error pattern in the peer-to-peer network. Our method could facilitate the rapid detection of mutations associated with various cancers.https://doi.org/10.1038/s42003-025-08537-3
spellingShingle Hyeonseob Lim
Soyeong Jun
Taehoon Kim
Ji Hyun Lee
Duhee Bang
Correcting errors in PCR-derived libraries for rare allele detection by reconstructing parental and daughter strand information
Communications Biology
title Correcting errors in PCR-derived libraries for rare allele detection by reconstructing parental and daughter strand information
title_full Correcting errors in PCR-derived libraries for rare allele detection by reconstructing parental and daughter strand information
title_fullStr Correcting errors in PCR-derived libraries for rare allele detection by reconstructing parental and daughter strand information
title_full_unstemmed Correcting errors in PCR-derived libraries for rare allele detection by reconstructing parental and daughter strand information
title_short Correcting errors in PCR-derived libraries for rare allele detection by reconstructing parental and daughter strand information
title_sort correcting errors in pcr derived libraries for rare allele detection by reconstructing parental and daughter strand information
url https://doi.org/10.1038/s42003-025-08537-3
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AT soyeongjun correctingerrorsinpcrderivedlibrariesforrarealleledetectionbyreconstructingparentalanddaughterstrandinformation
AT taehoonkim correctingerrorsinpcrderivedlibrariesforrarealleledetectionbyreconstructingparentalanddaughterstrandinformation
AT jihyunlee correctingerrorsinpcrderivedlibrariesforrarealleledetectionbyreconstructingparentalanddaughterstrandinformation
AT duheebang correctingerrorsinpcrderivedlibrariesforrarealleledetectionbyreconstructingparentalanddaughterstrandinformation