一种改良的植物DNA提取方法

植物组织中含有大量多糖、多酚、酯类等次生代谢产物, 要从中提取高质量的DNA比较困难。针对这一情况, 该文提出一种改良CTAB植物DNA提取方法(mCTAB), 并以10种常见植物为实验材料, 与4种常用的植物DNA提取试剂盒作对比。结果表明, mCTAB法提取的DNA产率高且质量好, PCR扩增成功率也较高, 而提取成本显著低于DNA提取试剂盒, 可有效用于植物DNA条形码等研究的植物DNA提取。     

栗属植物基因组DNA的提取及RAPD、SSR分析

以中国板栗、茅栗和锥栗的叶片为材料,提取栗属的DNA。针对栗属植物富含多酚类等次生物质的特点对栗属DNA的提取方法进行了改良,采用CTAB-蛋白酶K法加重复抽提,去除栗属植物中的相关次生物质,获得了高质量的DNA。所提取的DNA完全满足PCR、RAPD、SSR等一些分子生物学实验要求。     

典型樱亚属植物基因组DNA改良提取方法研究

本实验在传统的CTAB法及试剂盒法的基础上,利用DNA提取缓冲液作为样品预处理液,配合其他操作步骤的优化,设计出针对樱亚属植物基因组DNA的新型提取方法,并对提取的DNA进行ISSR-PCR分子标记实验以检验其质量。实验结果表明,改进后的CTAB法及试剂盒法均能有效地去除样品中含有的多糖、色素、黄酮等杂质,而试剂盒法提取的DNA纯度和质量总体而言高于CTAB法。ISSR-PCR结果显示,两条引物对样品DNA均能进行有效扩增,并且扩增条带清晰,无明显降解。改良后的两种方法能高效高质量地提取樱亚属植物DNA,并且具有较高的通用性,可以运用于其他物种DNA的提取工作中。     

同时提取油茶中DNA和RNA的简便方法

介绍了一种以CTAB提取方法为基础,结合其它DNA和RNA提取方法,经反复实验建立的一种同时提取油茶DNA和RNA的简便方法。该方法能有效地去除植物组织中酚类和多糖等次生物质的影响,所得到的DNA和RNA纯度高,完整性好,可以用于进一步的如DNA分析,mRNA的分离,RT-PCR,构建cDNA文库和基因表达分析等实验。     

甘草属植物DNA提取方法研究

甘草属植物黄酮类、多糖等次生代谢产物高,严重影响了的DNA提取质量和产量。本研究通过对甘草属植物DNA不同提取材料和4种DNA提取方法的比较研究,筛选出一种适合甘草属植物的DNA提取方法,该方法可有效去除次生代谢产物对DNA的干扰,能较好的应用于RAPD扩增和遗传多样性分析。     

一种高效的植物DNA提取和PCR扩增体系建立

报道一种高效简易的植物DNA提取和PCR扩增体系。该体系仅需一步即可提取DNA,扩增时延长PCR预变性时间便能获得较好的目的基因扩增结果。本体系具有较高的实验稳定性和较好的物种适用性,将大大提高植物分子生物学实验和基于分子标记辅助的植物育种实验效率,节省科研成本。     

松科植物基因组总DNA提取方法的比较

目的:研究对比了用不同方法提取红松、樟子松和红皮云杉等3种松科植物叶片基因组DNA的效果,以寻找适合提取松科植物叶片基因组DNA的方法。方法:分别比较了用传统的CTAB法、SDS法以及3种改良的CTAB法提取的上述3种松科植物叶片基因组DNA的电泳、纯度和酶切效果。结果:采用不同方法提取的3种松科植物叶片基因组DNA的质量差异较大。结论:常规CTAB法和SDS法无法提取出高质量松科植物基因组DNA,而改良后的CTAB法的提取效果较好。     

一种改良的植物基因组DNA通用提取方法

高质量的基因组DNA是分子生物学研究的基础,而从富含糖类和次生代谢物且异质性强的植物材料中分离DNA相对困难。本方法在CTAB法和商业DNA提取试剂盒的基础上,在裂解细胞之前,对植物材料进行预处理．去除干扰DNA提取的代谢物,并在后续步骤中进行了一些优化。该方法适于多种不同的植物种类,所提取的基因组DNA质量较好,能满足下一步基因操作的要求,是一种通用的植物基因组DNA提取方法。     

红豆杉属植物三种不同总DNA提取方法的分析比较

红豆杉属植物均为濒危物种,也是国家一级保护植物.以红豆杉属植物叶片为材料,利用三种不同的DNA提取方法提取总DNA,用分光光度计和琼脂糖凝胶电泳方法检测所得总DNA的得率和纯度,用PCR扩增的方法检测所得总DNA的质量,并对三种不同提取方法的结果进行了比较分析.结果表明:CTAB法提取的DNA纯度和得率均较高,可直接用...     

微量植物样本的DNA提取方法研究

为建立一种适于法庭科学实践的植物物证DNA提取优化方法,以期获得高质量的适于PCR分析的模板DNA.用8种方法从不同植物的干叶片中提取DNA,利用线粒体DNA非编码区的PCR扩增结果分析评价提取DNA的质量.结果表明8种DNA提取方法所提取的DNA都可以获得线粒体DNA非编码区的PCR扩增产物,对照紫外波长扫描结果显示,以改进的CTAB方法制备的模板DNA纯度最高,可达到进口试剂盒同等制备精度,OD260/280稳定在1.7～1.9之间.因此改进的CTAB方法适用于微量植物样本的DNA提取,可应用于法庭科学实践.     

一种快速微量提取植物叶片DNA的方法

介绍了一种快速提取微量DNA的方法。该方法简单易行,无需任何特殊设备,所需样品量少。提取的DNA纯度高,D260nm/D280nm在1．9-2．1之间,可满足RAPD、SSR、转基因植株的PCR检测等以PCR扩增为基础的实验需要。     

一种快速握取植物总DNA的良好材料与方法

本文介绍以种子植物幼嫩胚乳为材料,用改良的氯仿－异戊醇快速提取方法提取植物总ＤＮＡ。这种材料和方法,细胞磨碎容易,ＤＮＡ收取量大,操作简便,适用于遗传操作及教学实验等方面的ＤＮＡ快速制备。     

DNA barcode ITS2 coupled with high resolution melting (HRM) analysis for taxonomic identification of Sideritis species growing in Greece

Identification of genotypes in Sideritis is complicated owing to the morphological similarity and common occurrence of natural hybridisation within Sideritis species. Species- and genotype-specific DNA markers are very useful for plant identification, breeding and preservation programs. Herein, a real-time polymerase chain reaction (PCR) of ITS2 barcode region coupled with high resolution melting-curve (HRM) analysis was evaluated for an accurate, rapid and sensitive tool for species identification focusing on seven Sideritis species growing in Greece. The HRM assay developed in this study is a rapid and straightforward method for the identification and discrimination of the investigated Sideritis species. This assay is simple compared to other genotyping methods as it does not require DNA sequencing or post-PCR processing. Therefore, this method offers a new alternative for rapid detection of Sideritis species.     

A Simple and Non-destructive Method of Direct-PCR for Plant Systems

To date, PCR is a fundamental tool for most of the research concerning plant diversity analysis, marker-assisted selection, genetic purity testing, disease diagnostics, and transgene analysis. In all of these analyses, good-quality DNA serves as a template for amplification of target sequences. Extraction of good-quality DNA requires many steps, making the whole process time consuming, tedious, labor intensive, and expensive due to costlier and toxic chemicals. To overcome these preparatory steps from PCR-based DNA amplification, we have developed a direct-PCR amplification method for plants without isolating DNA. The method is unique and beneficial over some previously described methods of direct-PCR which fail due to inefficient amplification of target DNA in the presence of PCR inhibitors and crop specificity. Moreover, such methods are non-specific and, being destructive, cannot be replicated; one cannot completely rely on them due to lack of reproducibility. This method was streamlined from our earlier observation that alcohol-desiccated tissues maintain intact DNA for a long time. This method is specific, rapid, and, being non-destructive, allows replication, giving advantages over existing methods. The method was tested over a wide range of plant species and found very effective and quick in generating data. The method was successfully used to test the genetic purity of pearl millet hybrid (RHB-127) and its restorer (RIB 3135-18) and CMS line (ICMA 93333A). Our method is especially important for developing inexpensive and high-throughput non-invasive genetic analyses.     

Rapid isolation of high molecular weight plant DNA.

A method is presented for the rapid isolation of high molecular weight plant DNA (50,000 base pairs or more in length) which is free of contaminants which interfere with complete digestion by restriction endonucleases. The procedure yields total cellular DNA (i.e. nuclear, chloroplast, and mitochondrial DNA). The technique is ideal for the rapid isolation of small amounts of DNA from many different species and is also useful for large scale isolations.     

一种适于转基因水稻PCR检测的微量DNA快速提取法

对已报道的小麦基因组DNA快速提取方法的部分步骤进行了简化,在水稻上进行了尝试。结果表明,简化法提取的水稻基因组DNA完整性好,PCR扩增效果与试剂盒提取法无明显的差异,结果稳定可靠;而且整个提取过程操作简单、花费时间少,样品用量少,仅需5-10mg,适用于大规模转基因水稻的PCR检测。     

转基因植物快速检测方法的研究

本试验对转基因植物检测中的DNA提取和PCR扩增程序作了改进。经试验,本研究建立的DNA快速提取法与目前广泛使用的CTAB法相比更为简便,快速和经济,提取的DNA质量主扩增效果无明显差异,可用于多种转基因植物,多种植物组织的DNA提取,利用复合PCR法可在同一反应管中同步检测35N,NOS及CP4－EPSPS基因,明显提高了检测效率。应用本试验建立的DNA快速提取－复合PCR扩增－银染检测技术可在6小时内得出结果,达到了快速,简便,灵敏,可靠的检测目的。     

Differential Giemsa staining in plants : III. DNA base composition

DNAs isolated from three cultivars of Tulipa displaying a range of constitutive heterochromatin (<10% to 40%), showed very little or no difference in DNA base composition as determined from buoyant densities and thermal transition profiles. Four possible explanations for the interactions of the Giemsa dye and the chromatin are discussed with reference to the mechanism of Giemsa banding. A method for the rapid isolation of higher plant DNAs is described.     

A Rapid DNA Extraction Method for RFLP and PCR Analysis from a Single Dry Seed

A single-seed DNA extraction method was developed for rapid identification of plant genotype. The method was applied to 12 plant species, including the oil seeds sesame and soybean. The results were comparable to those obtained for oil-less seeds such as rice. This method will be useful for genotypic selection which requires rapid screening of large populations. It can also be used to identify varietal purity of seed stocks by PCR and RFLP analysis. The method includes two major steps, (i) treatment by proteinase K in an SDS extraction buffer, and (ii) grinding of a single half seed in the buffer after incubation. About 1.5–2 µg of DNA per half seed (the endosperm part) of rice was obtained and more than 200 half seed samples could be handled by one person in a day. The DNA could be used for fingerprinting and detection of target genes in a transgenic plant by PCR. The amplified PCR products from the half seed DNA exhibited the same banding patterns as those from leaf DNA. Yield and quality of DNA extracted from half seeds of rice was also sufficient for RFLP analysis. The remnant half seeds containing the embryo can be maintained for later germination of selected genotypes.     

Rapid isolation of terminal sequences from cloned plant DNA fragments

The isolation of DNA clone termini is an important step in the development of DNA contigs utilized for a range of applications, including physical mapping, genetic map-based cloning, insertion mutagenesis cloning, and isolation of complete gene sequences. We describe a rapid PCR-based method for the isolation of vector-insert junctions, or insert terminal sequences, of cloned plant DNA fragments. PCR amplification is performed using a vector-specific primer and a nonspecific primer, originally designed for use in animal systems, containing degenerative bases that we have shown can also anneal to plant insert DNA. Using this method we have successfully isolated end-terminal sequences from plant genomic clones harbored in YAC, BAC, and bacteriophage λ vectors. Termini of genomic clones from both tomato andArabidopsis were isolated demonstrating the utility of this technique among a range of plant species.