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四引物扩增受阻突变系统(T-ARMS)。

Tetra-Primer Amplification Refractory Mutation System (T-ARMS).

机构信息

Department of Plant Breeding and Genetics, Bihar Agricultural College, Bihar Agricultural University, Sabour, Bhagalpur, Bihar, India.

出版信息

Methods Mol Biol. 2023;2638:315-325. doi: 10.1007/978-1-0716-3024-2_22.

Abstract

Single-nucleotide polymorphisms (SNPs), the most abundant genetic variation in the population, have become the molecular marker of choice. Generally, the efficient detection of SNPs requires specialized costly equipment. Although there are a few strategies for detecting SNPs through polymerase chain reaction, followed by restriction enzyme digestion and agarose gel electrophoresis, these methods are time-consuming and might be less diagnostic. Interestingly, the tetra primer amplification refractory mutation system (T-ARMS) strategy utilizes a pair of allele-specific primers in a single PCR for the diagnostic detection of SNPs in a codominant manner through standard agarose gel electrophoresis. The simplicity and robustness of the strategy have inspired the researchers to adopt this low-cost method of SNP detection in different crop plants. Here, we have described the principle, methods, and conditions for the T-ARMS strategy. The described methodology starts from the isolation of genomic DNA and ends with the post-PCR analysis of refractory amplicons in standard agarose gel electrophoresis. The limitations and future perspectives are also discussed. Taken together, T-ARMS evolves as a method of choice for low-cost SNP detection in plants.

摘要

单核苷酸多态性(SNPs)是人群中最丰富的遗传变异,已成为首选的分子标记。通常,SNP 的高效检测需要专门的昂贵设备。虽然有一些通过聚合酶链反应(PCR)检测 SNPs 的策略,随后是限制性内切酶消化和琼脂糖凝胶电泳,但这些方法耗时且可能诊断性较差。有趣的是,四引物扩增受阻突变系统(T-ARMS)策略利用一对等位基因特异性引物在单个 PCR 中以共显性方式进行 SNP 的诊断检测,通过标准琼脂糖凝胶电泳进行。该策略的简单性和稳健性激发了研究人员在不同作物植物中采用这种低成本 SNP 检测方法。在这里,我们描述了 T-ARMS 策略的原理、方法和条件。所描述的方法学从基因组 DNA 的分离开始,最后在标准琼脂糖凝胶电泳中对不可扩增的扩增子进行 PCR 后分析。还讨论了该方法的局限性和未来展望。总之,T-ARMS 作为一种用于植物中低成本 SNP 检测的方法,是一种不错的选择。

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