A high-throughput protocol for deamination of long single-stranded DNA and oligo pools containing complex sequences

Cytidine deaminases as DNA mutators play important roles in immunity and genome stability. Here, we present a high-throughput protocol for deamination of long single-stranded (ss) DNA or oligo pools containing complex sequences. We describe steps for the preparation of both enzyme (activation-induce...

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Detalles Bibliográficos
Autores principales: Wang, Yanyan, Zhang, Senxin, Zheng, Xiaoqi, Yeap, Leng-Siew, Meng, Fei-Long
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10522977/
https://www.ncbi.nlm.nih.gov/pubmed/37742176
http://dx.doi.org/10.1016/j.xpro.2023.102602
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author Wang, Yanyan
Zhang, Senxin
Zheng, Xiaoqi
Yeap, Leng-Siew
Meng, Fei-Long
author_facet Wang, Yanyan
Zhang, Senxin
Zheng, Xiaoqi
Yeap, Leng-Siew
Meng, Fei-Long
author_sort Wang, Yanyan
collection PubMed
description Cytidine deaminases as DNA mutators play important roles in immunity and genome stability. Here, we present a high-throughput protocol for deamination of long single-stranded (ss) DNA or oligo pools containing complex sequences. We describe steps for the preparation of both enzyme (activation-induced deaminase, AID) and ssDNA substrates, the deamination reaction, uracil-friendly amplification, and data analysis. This assay can be used to determine the intrinsic mutation profile of a single antibody gene or a pool of selected regions on genomic DNA. For complete details on the use and execution of this protocol, please refer to Wang et al. (2023).(1)
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spelling pubmed-105229772023-09-28 A high-throughput protocol for deamination of long single-stranded DNA and oligo pools containing complex sequences Wang, Yanyan Zhang, Senxin Zheng, Xiaoqi Yeap, Leng-Siew Meng, Fei-Long STAR Protoc Protocol Cytidine deaminases as DNA mutators play important roles in immunity and genome stability. Here, we present a high-throughput protocol for deamination of long single-stranded (ss) DNA or oligo pools containing complex sequences. We describe steps for the preparation of both enzyme (activation-induced deaminase, AID) and ssDNA substrates, the deamination reaction, uracil-friendly amplification, and data analysis. This assay can be used to determine the intrinsic mutation profile of a single antibody gene or a pool of selected regions on genomic DNA. For complete details on the use and execution of this protocol, please refer to Wang et al. (2023).(1) Elsevier 2023-09-23 /pmc/articles/PMC10522977/ /pubmed/37742176 http://dx.doi.org/10.1016/j.xpro.2023.102602 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Wang, Yanyan
Zhang, Senxin
Zheng, Xiaoqi
Yeap, Leng-Siew
Meng, Fei-Long
A high-throughput protocol for deamination of long single-stranded DNA and oligo pools containing complex sequences
title A high-throughput protocol for deamination of long single-stranded DNA and oligo pools containing complex sequences
title_full A high-throughput protocol for deamination of long single-stranded DNA and oligo pools containing complex sequences
title_fullStr A high-throughput protocol for deamination of long single-stranded DNA and oligo pools containing complex sequences
title_full_unstemmed A high-throughput protocol for deamination of long single-stranded DNA and oligo pools containing complex sequences
title_short A high-throughput protocol for deamination of long single-stranded DNA and oligo pools containing complex sequences
title_sort high-throughput protocol for deamination of long single-stranded dna and oligo pools containing complex sequences
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10522977/
https://www.ncbi.nlm.nih.gov/pubmed/37742176
http://dx.doi.org/10.1016/j.xpro.2023.102602
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