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通过对DNAdvance试剂盒进行析因设计来降低DNA提取成本。

Reducing DNA extraction costs through factorial design for the DNAdvance Kit.

作者信息

Neal Carson J, Zbinden Zachery D, Douglas Michael E, Douglas Marlis R

机构信息

Department of Biological Sciences, University of Arkansas, Fayetteville, AR, USA.

Appalachian Laboratory, University of Maryland Center for Environmental Science, Frostburg, MD, USA.

出版信息

BMC Res Notes. 2024 Dec 30;17(1):397. doi: 10.1186/s13104-024-07063-5.

Abstract

OBJECTIVE

Extracting DNA is essential in wildlife genetic studies, and numerous methods are available. However, the process is costly and time-consuming for non-model organisms, including most wildlife species. Therefore, we optimized a cost-efficient protocol to extract DNA from the muscle tissue of White-tailed Deer using the DNAdvance kit (Beckman Coulter), a magnetic-bead-based approach. We devised a 3 × 3 factorial design using combinations of tissue mass (10 mg, 50 mg, or 100 mg) and reaction volume (25%, 33%, and 50% of the manufacturer's recommended volumes). DNA was extracted for N = 81 tissue sub-samples (9 replicates/treatment).

RESULTS

Our target yield was 500 ng of genomic DNA per sample, sufficient for population genetic assessments. A combination of 50 mg tissue and 25% reaction volume yielded enough DNA at the lowest cost. The factorial design revealed that varying tissue mass and reagent volume significantly affected extracted DNA yield. Our study demonstrates that sufficient DNA can be extracted at 75% lower costs than the manufacturer's standard protocol. Other researchers can directly use our modified DNAdvance protocol to perform cost-effective DNA extractions.

摘要

目的

提取DNA在野生动物遗传研究中至关重要,且有多种方法可用。然而,对于非模式生物,包括大多数野生动物物种而言,该过程成本高昂且耗时。因此,我们优化了一种经济高效的方案,使用基于磁珠法的DNAdvance试剂盒(贝克曼库尔特公司)从白尾鹿的肌肉组织中提取DNA。我们采用组织质量(10毫克、50毫克或100毫克)和反应体积(制造商推荐体积的25%、33%和50%)的组合设计了一个3×3析因设计。对N = 81个组织子样本(每个处理9个重复)进行了DNA提取。

结果

我们的目标产量是每个样本500纳克基因组DNA,足以进行群体遗传评估。50毫克组织和25%反应体积的组合以最低成本产生了足够的DNA。析因设计表明,不同的组织质量和试剂体积显著影响提取的DNA产量。我们的研究表明,与制造商的标准方案相比,可将成本降低75%提取到足够的DNA。其他研究人员可以直接使用我们改良的DNAdvance方案进行经济高效的DNA提取。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81cc/11684086/121ef5effa17/13104_2024_7063_Fig1_HTML.jpg

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