一种改良的SDS法用于从生玉米和烤大豆中提取高质量DNA

IF 0.4 Q4 VETERINARY SCIENCES
Arita Sabriu-Haxhijaha, G. Ilievska, V. Stojkovski, K. Blagoevska
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引用次数: 2

摘要

摘要随着全球转基因作物种植面积的扩大,非转基因品种被转基因产品污染的可能性也在增加。基于dna的方法作为准确、高效、可靠的检测原料或深加工食品中转基因物质的优选方法。采用合适、高效的DNA提取方案分离高质量的DNA是获得精确DNA扩增结果的关键。在本研究中,我们对已知的基于十二烷基硫酸钠(SDS)的DNA提取方法在孵育时间、DNA颗粒洗涤和添加有机溶剂提取等方面进行了改进,以提高DNA质量并降低成本。以生玉米粒和烤大豆种子为样品。DNA提取遵循三个方案,爱德华兹方案的修改。原料玉米样品中使用的洗涤剂类型对提取DNA的产量和纯度没有显著影响,而在烤大豆样品中,2% (w/v)的SDS裂解缓冲液的DNA产量最高。另外的孵育步骤使玉米的DNA得率提高了121%,而大豆样品的DNA纯度则是通过有机溶剂萃取得到的。DNA完整性的电泳测定表明,烤大豆中存在不同程度的DNA涂布。相反,所有对生玉米粒的提取都产生了高分子量的DNA。因此,与商业试剂盒相比,我们的内部DNA提取方案同样高效,但更具成本效益,可用于生玉米,而用于烤大豆的方案需要进一步改进。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A Modified SDS – Based Method Applied for Extraction of High-Quality DNA from Raw Corn and Roasted Soybean
Abstract The probability of contamination of non-transgenic varieties with genetically modified (GM) products increase as a result of global expansion of areas sown with transgenic crops. DNA-based methods as accurate, efficient and reliable methods are preferable for detection of GM material in raw or highly processed foods. Isolation of high quality DNA with a suitable and efficient DNA extraction protocol is crucial for getting precise results in DNA amplification. In this study, we performed modifications of previously known Sodium dodecyl sulfate (SDS)-based DNA extraction method regarding the incubation period, DNA pellet washing and addition of organic solvent extraction, to improve DNA quality and to reduce costs. Raw corn kernels and roasted soybean seed were used as samples. DNA was extracted following three protocols, modifications of Edwards protocol. The type of detergent used in raw corn sample did not cause significant effects on extracted DNA yield and purity, while in roasted soybean samples the 2% (w/v) SDS lysis buffer gave the highest DNA yield. The additional incubation step raised the DNA yield from raw corn for 121%, while the purest DNA from soybean sample was obtained using organic solvent extraction. Electrophoretic determination of DNA integrity showed varying degree of DNA smearing from roasted soybean. Contrary, all extraction protocols used on raw corn kernels produced a high molecular weight DNA. Thus, our in-house DNA extraction protocol is as efficient but more cost effective compared to commercial kits and can be used for raw corn, while the protocol for roasted soybean needs further improvement.
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来源期刊
Macedonian Veterinary Review
Macedonian Veterinary Review Veterinary-Veterinary (all)
CiteScore
1.00
自引率
0.00%
发文量
20
审稿时长
12 weeks
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