数字农科院2.0

Two-step method for rapid isolation of genomic DNA and validation of R81T insecticide resistance mutation in Myzus persicae

文献类型： 外文期刊

作者： Muhammad Umair Sial;Tahir Farooq;Luaay Kahtan Khalaf;Saqib Rahman;Muhammad Asad;Bilal Ahamad Paray

作者机构：

关键词： DNA;Myzus persicae;Phenol:chloroform;Purity;R81T;Salt out;Yield

期刊名称： Saudi Journal of Biological Sciences

ISSN： 1319-562X

年卷期： 2023 年 30 卷 11 期

页码：

收录情况： SCIE（2023版）

摘要： Isolation and amplification of nucleic acid (DNA) is considered a vital and potent instrument in molecular biological research. However, its functioning outside of a laboratory setting is difficult because of complex procedures that demand expert personnel and expensive equipment in addition to the fulfillment of several additional requirements. DNA isolation from minute insects is sometimes difficult, making diagnostic and genotyping procedures problematic. Thus, the current work offers a high-throughput, cost-effective, straightforward, and faster approach for isolating DNA from the aphid Myzus persicae. Intriguingly, two-step DNA extraction process yielded a high yield of extremely pure genomic DNA and required only 10 s to complete. PCR investigation aiming at amplifying the non-synonymous R81T region on the loop D site of the nAChR gene of M. persicae was subsequently utilized to successfully validate the recovered DNA. Moreover, the proposed method was compared in terms of yield and purity with conventionally used DNA isolation methods including, phenol:chloroform, salt out, and commercially available kits. In conclusion, this newly developed method would enable researchers to quickly process many biological samples used to analyze genetic diversity, mutant screening, and large spectrum diagnosis both in laboratory and field conditions.

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