Xiao Cheng, Xiaoling Hong, M. Khayatnezhad, F. Ullah
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Our results showed significant differences in DNA contents between these five methods. The quantity and quality of extracted genomic DNA were checked by the spectrophotometer, Nano-Drop and and agarose gel electrophoresis analysis. Finally, a PCR-based method was also applied to verify the amplification efficiency for two molecular markers (ITS and ISSR).. In the present study, the genetic diversity of 96 Tamarix individuals species and 8 populations were studied using 10 ISSR markerswhile for nrDNA ITS 8 species samples were used. The method of Nalini et al., provided best results (207 ng/μL) in terms of quantity and quality ofDNA. Our results proposed that this method could be effective for plants with the same polysaccharides, proteins and polyphenols components. The advantage of this method is simple and fast as it does not involve time consuming steps such as incubation at higher temperatures, and also do not requires expensive chemicals such as proteinase K, liquid nitrogen. ,. 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引用次数: 18
摘要
柽柳属约有54种,主要生长在沙漠和半沙漠的盐碱地。这种属的化学特征是单宁,类黄酮,花青素和精油的存在干扰纯基因组DNA的提取。因此,有必要优化提取方案,以尽量减少这些化合物的影响到最低水平。本研究采用Kit (DNP TM Kit)、Murray and Thompson、Sahu et al.、Nalini et al.和Bi et al.的CTAB DNA提取方法提取柽柳物种的DNA,比较了五种不同方法提取柽柳物种基因组总DNA的效率,发现提取的DNA含量和质量存在显著差异。我们的结果显示,这五种方法的DNA含量有显著差异。采用分光光度计、纳米滴法和琼脂糖凝胶电泳法检测提取的基因组DNA的数量和质量。最后,采用pcr方法验证了ITS和ISSR两种分子标记的扩增效率。本研究利用10个ISSR标记对柽柳96个种和8个居群的遗传多样性进行了研究,并利用8个种样本进行了nrDNA ITS分析。以Nalini等人的方法(207 ng/μL)测定dna的数量和质量最好。结果表明,该方法对具有相同多糖、蛋白质和多酚成分的植物是有效的。这种方法的优点是简单和快速,因为它不涉及耗时的步骤,如在高温下孵育,也不需要昂贵的化学品,如蛋白酶K,液氮。,。该方法获得高质量基因组DNA的成功已在柽柳种群中得到证明,并对该方法的可靠性进行了讨论。
Genetic diversity and comparative study of genomic DNA extraction protocols in Tamarix L. species
The genus Tamarix consists of about 54 species that mainly grow in saline areas of deserts and semi-deserts. This genus is chemically characterized by the presence of tannins, flavonoids, anthocyanins and essential oils which interfere with the extraction of pure genomic DNA. Thus it is necessary to optimize extraction protocols to minimize the influence of these compounds to the lowest level. The present study compares the efficiency of five different approaches to extract total genomic DNA in Tamarix species, showing significant differences in the extracted DNA contents and quality,by using Kit (DNP TM Kit), CTAB DNA extraction method by Murray and Thompson, Sahu et al., Nalini et al. and Bi et al., for the extraction of DNA from Tamarix species. Our results showed significant differences in DNA contents between these five methods. The quantity and quality of extracted genomic DNA were checked by the spectrophotometer, Nano-Drop and and agarose gel electrophoresis analysis. Finally, a PCR-based method was also applied to verify the amplification efficiency for two molecular markers (ITS and ISSR).. In the present study, the genetic diversity of 96 Tamarix individuals species and 8 populations were studied using 10 ISSR markerswhile for nrDNA ITS 8 species samples were used. The method of Nalini et al., provided best results (207 ng/μL) in terms of quantity and quality ofDNA. Our results proposed that this method could be effective for plants with the same polysaccharides, proteins and polyphenols components. The advantage of this method is simple and fast as it does not involve time consuming steps such as incubation at higher temperatures, and also do not requires expensive chemicals such as proteinase K, liquid nitrogen. ,. The success of this method in obtaining high-quality genomic DNA has been demonstrated in the Tamarix species group and the reliability of this method has been discussed.
期刊介绍:
Caryologia is devoted to the publication of original papers, and occasionally of reviews, about plant, animal and human karyological, cytological, cytogenetic, embryological and ultrastructural studies. Articles about the structure, the organization and the biological events relating to DNA and chromatin organization in eukaryotic cells are considered. Caryologia has a strong tradition in plant and animal cytosystematics and in cytotoxicology. Bioinformatics articles may be considered, but only if they have an emphasis on the relationship between the nucleus and cytoplasm and/or the structural organization of the eukaryotic cell.