T7 DNA 聚合酶处理可提高双链和单链 DNA 病毒的定量测序能力

Peer community journal Pub Date : 2024-07-11 DOI:10.24072/pcjournal.437

Maud Billaud, Ilias Theodorou, Quentin Lamy-Besnier, Shiraz A. Shah, François Lecointe, Luisa De Sordi, Marianne De Paepe, Marie-Agnès Petit

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摘要

大量微生物组和富集病毒组的霰弹枪测序只能揭示特定样本中的双链 DNA（dsDNA）含量，除非进行特殊处理。然而，病毒基因组通常由环状单链 DNA（ssDNA）分子组成。使用多重位移扩增（MDA）法对 DNA 进行预处理和扩增，可将 ssDNA 转化为 dsDNA，但这一过程会导致这些环状 ssDNA 基因组的过度呈现。最新的替代方法允许绕过扩增

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T7 DNA polymerase treatment improves quantitative sequencing of both double-stranded and single-stranded DNA viruses

Bulk microbiome, as well as virome-enriched shotgun sequencing only reveals the double-stranded DNA (dsDNA) content of a given sample, unless speciﬁc treatments are applied. However, genomes of viruses often consist of a circular single-stranded DNA (ssDNA) molecule. Pre-treatment and ampliﬁcation of DNA using the multiple displacement ampliﬁca-tion (MDA) method enables conversion of ssDNA to dsDNA, but this process can lead to over-representation of these circular ssDNA genomes. A more recent alternative permits to bypass the ampliﬁcation

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Peer community journal

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