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苎麻总DNA提取的CTAB法优化方案 总被引:17,自引:0,他引:17
根据苎麻组织自身含有较多的本分类,粘状物及黄酮类等次生代谢物质的特点,对CTAB法进行优化后用于提取苎麻组织总DNA。在研磨过程中加入抗氧化剂PVP,提取缓冲液加入终浓度6%的PVP和4%的β-巯基乙醇,可防止整个提取过程中溶液变褐;在第2次用氯仿/异戊醇抽提时,加入10%的CTAB和4%的NaCl高盐溶液,以除去多糖。经几种改良方案提取总DNA的ISSR-PCR扩增结果验证,以同时加入抗氧化剂和高盐溶液的CTAB-4法效果最好。 相似文献
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一种改良的CTAB法提取产多糖真菌DNA 总被引:5,自引:0,他引:5
真菌胞外多糖由于其高吸附高粘稠特点,是困扰从胞外多糖产生菌分离高纯度DNA的难点之一。本文以生产硬葡聚糖的齐整小核菌生产菌为代表,采用改良的CTAB法获得了高质量的基因组DNA。通过分层隔离等培养方法的优化降低硬葡聚糖的产生,并在传统CTAB法的基础上,用高浓度的醋酸钾和无水乙醇共同作用初步沉淀多糖,再用CTAB/NaCl溶液再次去除多糖。相比于商业的DNA提取试剂盒和传统的CTAB法,该方法得到的基因组DNA产率大幅提高,纯度较好,可充分排除胞外多糖的干扰,为各典型产胞外多糖的真菌DNA提取提供重要的参考。提取的基因组DNA可用于基因组文库构建、PCR等分子生物学实验。 相似文献
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应用改良CTAB法提取树莓DNA最佳条件的探索 总被引:1,自引:0,他引:1
目的:改良CTAB法提取三种树莓叶DNA最佳条件的探索及检测DNA的完整度。方法:通过改良CTAB法提取树莓叶三个品种的基因组DNA。结果:经检测,所提样品OD260/OD280值在1.6~1.9之间,得率为198~396ng/μl,电泳条带较清晰,无明显拖尾现象;提取最佳条件为水浴时间90min、CTAB抽提液浓度3%时,发现DNA完整性较好、纯度较高,可以满足相关的分子生物学研究需要。结论:该研究可以为树莓叶分子生物学的后续研究的开展奠定实验基础。 相似文献
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元宝枫叶片总DNA提取方法的优化 总被引:3,自引:0,他引:3
由于元宝枫叶片细胞含有大量的多糖、单宁、色素和酚类物质,严重影响基因组DNA的提取,在提取元宝枫叶片总DNA的过程中,对常用的CTAB法进行了大胆的革新.结果表明,采用细胞核裂解前加入不溶性聚乙烯吡咯烷酮(PVP)和将加入RNase A消化RNA的步骤改在最后进行等技术,可以从元宝枫叶片快速提取高质量DNA,该研究为从富含多糖、单宁、色素和酚类物质的植物中提取基因组DNA,提供了可行的分析方法. 相似文献
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应用CTAB法对砂梨品种DNA提取效果的研究 总被引:4,自引:0,他引:4
为了获得质量较好的DNA,我们采用CTAB法对11个砂梨品种的叶片DNA提取情况进行了研究,发现参试样品中一些样品的蛋白质含量较多,而另外一些样品的多糖、酚类物质含量较高。对于蛋。白质含量较多的这类材料,适当增加24:1的处理次数能够使蛋白质沉淀下来;对于多糖、酚类物质含量较高的材料,用5mol/L的NaCl和3mol/L的NaAc等处理能够有效的降低多糖含量。另外,对研磨样品的加入量和PVP的加入时间进行了对比,最后用双酶切、电脉检测等不同的分析方法对所得到的DNA提取物的浓度和纯度进行检测,发现加入样品量在0.3g左右、在研磨后的粗提液中加入PVP,所得到的DNA质量较好. 相似文献
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CTAB法提取野野村菌基因组DNA 总被引:4,自引:0,他引:4
针对用常规方法难以提取野野村菌基因组DNA的问题,通过选用添加甘氨酸的不同培养基和不同培养时间获得的菌丝体,采用液氮研磨结合CTAB法提取野野村菌DNA,电泳检测及计算OD260/OD280值。结果表明,在添加0.3%甘氨酸的麦芽汁-酵母膏(YE)培养基中振荡培养培养3d的菌丝体适合于DNA提取,用CTAB法获得的基因组DNA,长度约为20kb,且OD260/OD280在1.8左右,达到基因组DNA-DNA杂交的要求。 相似文献
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高盐沉淀CTAB法提取温室菊花基因组DNA 总被引:4,自引:0,他引:4
根据温室菊花植物组织富含多酚、多糖的具体特性,对CTAB法加以改进:在待沉淀液中加入1/2体积5 mol·L~NaCI.改进后的方法获得的DNA质量良好,电泳条带清晰,提取过程无明显的DNA降解,基本上排除了多酚物质的干扰.以提取的DNA为模板,用一对引物扩增菊花中18S基因,得到条带单一,大小与已知一致,说明获得的DNA可以进行PCR扩增,EcoR I 酶切基因组DNA图谱表明,提取的DNA能被限制性内切酶完全酶切,可以满足相关的分子生物学研究. 相似文献
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利用改良CTAB法快速小量提取微胚乳玉米基因组DNA 总被引:2,自引:0,他引:2
为了能快速小量提取微胚乳玉米基因组DNA,以微胚乳玉米幼叶为试材,采用改良CTAB法和传统CTAB法提取微胚乳玉米基因组DNA,并对所提取的DNA通过紫外分光光度计、琼脂糖凝胶电泳和PCR扩增等方法进行检测。两种方法所得基因组DNA的OD260/OD280在1.8~1.9之间,电泳条带清晰,无蛋白质和RNA污染,DNA无明显降解,其浓度和纯度都适合基因工程实验操作的条件。改良CTAB法与传统CTAB法相比更简便快捷,可实现大批量的不同样本基因组DNA的同时提取,提供了一种简便、快捷、有效和实用的微量提取微胚乳玉米基因组DNA方法,可满足以PCR为基础的分子生物学研究。 相似文献
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A (GA)n and (GT)n microsatellite‐enriched library was constructed and 23 nuclear simple sequence repeat (SSR) loci were characterized in the guava species (Psidium guajava L.). All SSR loci were found to be polymorphic after screening for diversity in different cultivars, and across‐taxa amplification tests showed the potential transferability of most SSR markers in three other Psidium species. First to be published for P. guajava, this new SSR resource will be a powerful tool for genetic studies of guava, including cultivars identification and linkage mapping, as well as potentially for interspecific genetic studies within the genus Psidium. 相似文献
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设计正交实验,采用超声波法从番石榴叶中提取总黄酮,综合各因素确定最佳提取条件为乙醇浓度40%,固料比(番石榴干叶:浸提剂)=1g:60mL,浸泡25h超声波处理时间50min,获得提取液总黄酮含量为9.9167%。采用牛津杯法做抑菌实验,结果表明番石榴叶和果实的水提、醇提粗提液对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌、阴沟肠杆菌、溶血葡萄球菌等均有很好的抑菌效果。叶和果实的总黄酮提取液的抑菌作用也很明显。番石榴有望作为药食同源产品和天然食品防腐剂进行开发利用。 相似文献
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提取得到高质量的DNA样品是进行分子生物学研究的必要前提。为了找到一种适用于提取涡虫基因组DNA的常规方法,我们以东亚三角头涡虫为材料,分别用改良的CTAB法、SDS法、SDS-蛋白酶K法对涡虫的基因组DNA进行了制备,并对3种方法制备的涡虫基因组DNA进行了检测与比较。根据比较结果,我们认为改良的CTAB法最适合于涡虫基因组DNA的快速制备,为涡虫的分子生物学研究打下了基础。 相似文献
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采用溶剂法提取番石榴Psidium guajava叶总黄酮,通过对溶剂浓度、提取温度、时间、料液比等因素进行正交试验,优化番石榴叶总黄酮的最佳提取工艺条件。结果表明,溶剂法提取番石榴叶总黄酮的最佳工艺参数为:80 ℃加热,60%乙醇,料液比1∶15,加热回流提取1 h,在此条件下总黄酮提取率达3.51%。番石榴叶总黄酮粗提物对茄子白绢病菌、甘蓝黑斑病菌、白菜炭疽病菌、黄瓜枯萎病菌的室内抑菌EC50分别为184、209、180、102 mg·mL-1。 相似文献
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Chemical Compositions,Antioxidant and Antimicrobial Activities of Essential Oils of Psidium guajava L. Leaves from Different Geographic Regions in China
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Hydrodistilled essential oils (EO) of Psidium guajava L. leaves from different regions in China were analyzed by GC and GC/MS. The samples from Guangdong Province displayed high EO yields (0.61 – 0.75%, v/w). A total of 50 components, representing over 98.00% of the EOs, were identified and semi‐quantitatived. The major constituents of EOs included β‐caryophyllene (17.17 – 31.38%), γ‐gurjunene (9.17 – 15.22%), τ‐cadinol (1.35 – 10.02%) and calamenene (2.13 – 7.80%). The terpenoids in all sample oils were dominated by sesquiterpenes hydrocarbons (70.18 – 84.35%), followed by oxygenated sesquiterpenes (9.89 – 22.19%). The similarities and differences among EOs from different samples were evaluated by hierarchical cluster analysis and principal component analysis methods. The IC50 values of EOs from different regions were between 18.52 – 33.72 mg/ml (DPPH) and 13.12 – 25.15 mg/ml (ABTS+). The FRAP value of EO from Guangdong Province was 7.34 – 9.13 mmol Vc/g DM, while the FRAP value of EO from Taiwan Province was 2.29 – 2.36 mmol Vc/g DM. The antimicrobial tests revealed that EO had a higher antimicrobial activity against all Gram‐positive bacteria and two fungi. Moreover, EO from P. guajava leaves of Guangdong Province showed the highest antimicrobial activity. These properties can be considered in the design of industrial products and for further application in the food, pharmaceutical and cosmetic industries. 相似文献
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Jiaokun Li Chenchen Wu Fang Li Runlan Yu Xueling Wu Li Shen 《Preparative biochemistry & biotechnology》2019,49(1):21-29
Psidium guajava leaves are rich in health-promoting flavonoids compounds. For better utilization of the resource, the ultrasound-assisted aqueous extraction was investigated using Box-Behnken design under response surface methodology. A high coefficient of determination (R2?=?97.8%) indicated good agreement between the experimental and predicted values of flavonoids yield. The optimal extraction parameters to obtain the highest total flavonoids yield were ultrasonic power of 407.41?W, extraction time of 35.15?min, and extraction temperature of 72.69?°C. The average extraction rate of flavonoids could reach 5.12% under the optimum conditions. Besides, HPLC analysis and field emission scanning electron microscopy indicated that the ultrasonic treatment did not change the main component of flavonoids during extraction process and the higher flavonoids content was attributed by the disruption of the cell walls of guava particles. Thus, the extraction method could be applied successfully for large-scale extraction of total flavonoids from guava leaves. 相似文献