A Robust Closed-Tube Method for Resolving tp53M214K Genotypes.
A Robust Closed-Tube Method for Resolving tp53M214K Genotypes.
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用于解析 tp53M214K 基因型的稳健闭管方法。
DOI：
10.1089/zeb.2023.0030
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发表时间：
2024
期刊：
影响因子：
2
通讯作者：
Kendall,GenevieveC
中科院分区：
文献类型：
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作者：
Silvius,KatherineM;Kendall,GenevieveC
tp53M214Kzebrafish 突变体是一个多功能平台，可用于模拟多种人类疾病。然而，目前针对该突变体的基因分型方法需要漫长的实际操作过程，例如限制性消化和外包桑格测序。为了解决这一缺陷，我们利用高分辨率熔解分析技术结合并行串联野生型掺入方法来开发能够区分 tp53M214Kzygosity 的强大基因分型方案。在这项研究中，我们详细描述了我们的方法。我们预计，我们的基因分型方案将有利于利用 tp53M214Kzebrafish 突变体的研究人员，通过提供比传统方法更短的周转时间、更少的人员参与和更高的通量提供可靠的结果，从而减轻基因分型的负担并最大限度地提高研究效率。
Thetp53M214Kzebrafish mutant is a versatile platform with which to model a diverse spectrum of human diseases. However, currently available genotyping methods for this mutant require lengthy hands-on processes such as restriction digests and outsourced Sanger sequencing. To address this deficiency, we leveraged high-resolution melting analysis technology in conjunction with a parallel, in-tandem wild-type spike-in approach to develop a robust genotyping protocol capable of discriminatingtp53M214Kzygosity. In this study, we describe our method in detail. We anticipate that our genotyping protocol will benefit researchers utilizing thetp53M214Kzebrafish mutant by offering reliable results with a shorter turnaround time, lower personnel involvement, and higher throughput than traditional methods, thereby decreasing the burden of genotyping and maximizing research efficiency.