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一种快速提取小麦叶片总RNA的方法 总被引:17,自引:0,他引:17
从植物组织中提取高质量的RNA是进行植物分子生物学研究的必要前提和关键.同种植物不同器官的组织由于组成分的差异,提取RNA的方法也存在不同的难点.在苯酚法和氯化锂沉淀法的基础上,改进并提出了一种适合小麦叶片总RNA的快速提取方法,消除了蛋白质、DNA、多糖、多酚等污染.该方法提取的小麦叶片总RNA,完整性好、纯度高,可用于RT-PCR、N orthern杂交、RACE等实验操作,而且简单经济、快速、实验结果稳定,重复性好,还适合富含多糖和脂质的植物组织总RNA的提取. 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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提取高质量人参RNA的方法研究 总被引:13,自引:0,他引:13
针对人参组织多酚、多糖类物质含量较高的特点,比较了改良的异硫氰酸胍法、改良的CTAB法和改良的Trizol法等3种不同的RNA提取方法.3种改良的方法均能从人参组织中提取到总RNA.其中改良的Trizol法能有效地抑制酚类物质和多糖对总RNA提取的影响,能从成熟的叶片中获得高质量、完整性好的总RNA,每克新鲜组织RNA产量在90~120!g之间,电泳分析,28SrRNA亮度约为18SrRNA的2倍,A260/A280介于1.8~2.0之间.用改良的Trizol法分离的RNA,已成功进行了RT-PCR及人参叶cDNA文库构建等研究. 相似文献
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为从富含多糖、酚类物质的丹参幼叶中提取高质量的核总DNA,比较了4种DNA提取方法对丹参叶片的提取效果,结果表明改良CTAB法提取的DNA溶液颜色为无色透明、A260/A230为2.137、A260/A280为1.816值最好,是提取丹参基因组DNA的有效方法. 相似文献
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干种子高质量总RNA的快速提取方法 总被引:1,自引:0,他引:1
高效快速提取高质量的种子RNA是种子分子生物学研究的基础。现有的提取方法难以高效快速地从种子中得到高质量的总RNA。本试验有机地将改进SDS法和异硫氰酸胍法相结合,采用改进的酸性SDS提取液、不溶性PVPP(聚乙烯聚吡咯烷酮)阻止酚类氧化、KAc去除多糖、异丙醇沉淀RNA,可以高效地从0.01~0.1g水稻、大豆、蚕豆、芸豆、花生等干种子中提取到高质量总RNA。此法提取的总RNA,能够满足分子生物学研究的要求,可以进行反转录和RT-PCR反应,用于基因表达研究,并为从具相似成分的其他物种干种子提取总RNA提供参考方法。 相似文献
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一种适合从柑橘果皮提取总RNA的方法 总被引:3,自引:1,他引:2
柑橘果皮由于富含果胶、酚类物质等干扰RNA分离的物质,较难提取到纯度高的RNA。本试验建立了一种适于从柑橘果皮提取RNA的方法,从脐橙和蕉柑两种柑橘的果皮提取总RNA,经凝胶电泳、紫外分光光度法检测所提RNA的品质。研究结果表明,该法所提RNA条带清晰、无降解。OD260/OD280接近2.0,具有较高的纯度。RT-PCR试验结果进一步表明,该法提取的RNA纯度高,完全能够用于后续的分子生物学研究。 相似文献
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Factors affecting the extraction of intact ribonucleic Acid from plant tissues containing interfering phenolic compounds 总被引:5,自引:0,他引:5 下载免费PDF全文
Using conventional methods it is impossible to extract RNA as uncomplexed intact molecules from the leaves of grapevines (Vitis vinifera L.) and from a number of woody perennial species that contain high levels of reactive phenolic compounds. A procedure involving the use of high concentrations of the chaotropic agent sodium perchlorate prevents the binding of phenolic compounds to RNA during extraction. Analyses of the phenolics present in plant tissues used in these experiments indicate that there is a poor correlation between the total phenolic content and the complexing of RNA. However, qualitative analyses suggest that proanthocyanidins are involved in the tanning of RNA during conventional extractions. 相似文献
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Laura Jaakola Anna Maria Pirttilä Minna Halonen Anja Hohtola 《Molecular biotechnology》2001,19(2):201-203
A simple and efficient method is described for isolating high quality RNA from bilberry fruit. The procedure is based on the
use of hexadecyltrimethyl ammonium bromide (CTAB), polyvinylpyrrolidone (PVP), and β-mercaptoethanol in an extraction buffer
in order to eliminate the polysaccharides and prevent the oxidation of phenolic compounds. This method is a modification of
the one described for pine trees, and yields high-quality RNA suitable for cDNA based methodologies. This method is applicable
for a variety of plant tissues. 相似文献
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C. Huang J. F. Picimbon H. Q. Li Z. Li Q. Liu W. Liu 《Russian Journal of Plant Physiology》2012,59(1):129-133
Most of conventional RNA extraction methods failed to extract highly pure and integral RNA from peanut seeds because peanut
seeds are extremely rich in lipids, proteins, polysaccharides, and phenolic compounds. Here, we describe a new method, named
Peanut Improved Modified RNA extraction method (PIMRNAext), using SDS, Tris-saturated phenol, NaCl, and sarkosyl during the
extraction process, which are particularly successful for total RNA extraction from lipid- and polysaccharide-rich materials.
The proposed PIMRNAext method is simple and fast. It requires only conventional reagents and is completed within 2 h. Using
PIMRNAext gives very good yields of high quality peanut RNA. This method is about ten times more efficient than conventional
methods, and the RNA produced by it is compatible with further molecular biology experiments, such as RT-PCR. We propose to
use the PIMRNAext method to extract RNA from peanuts and peanut-like plant species not only for RT-PCR, but also for most
molecular biology techniques that need copies of pure RNA, such as microarray or cDNA library construction. 相似文献
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Development of an Efficient Protocol of RNA Isolation from Recalcitrant Tree Tissues 总被引:1,自引:0,他引:1
Isolation of RNA from recalcitrant tree tissues has been problematic due to large amounts of secondary metabolites and interfering
compounds in their cells. We have developed an efficient RNA extraction method, which yielded high-quality RNA preparations
from tissues of the lychee tree. The method reported here utilized EDTA, LSS, and CTAB to successfully inhibit RNase activities.
It was found that a high ionic strength brought about by 2 M NaCl was necessary. In addition, secondary metabolites and other
interfering compounds were effectively removed using sodium borate and PVPP under a deoxidized condition. The quality of purified
RNA was tested by both RACE and Northern blotting analysis, ensuring that the RNA could be used for subsequent gene expression
analysis. This method has been successfully applied to purify RNA from 15 other plant species. In conclusion, the protocol
reported here is expected to have excellent applications for RNA isolation from recalcitrant plant tissues. 相似文献
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Kolosova N Miller B Ralph S Ellis BE Douglas C Ritland K Bohlmann J 《BioTechniques》2004,36(5):821-824
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Dongping Zha Haisheng Cai Xueling Zhang Qinggang He Liting Chen Chunqing Qiu Shufang Xia 《Phyton》2022,91(10):2297-2311
The lotus (Nelumbo nucifera Gaertn.) is an aquatic plant that grows in shallow water and has long been cultivated
in South China. It can improve the incomes of farmers and plays an important role in alleviating poverty in rural
China. However, a modern method is required to accurately estimate the area of lotus fields. Lotus has spectral
characteristics similar to those of rice, grassland, and shrubs. The features surrounding areas where it is grown are
complex, small, and fragmented. Few studies have examined the remote sensing extraction of lotus fields, and
automatic extraction and mapping are still challenging methods. Here, we compared the spectral characteristics
of lotus fields and other ground objects and devised a remote sensing method for the rapid extraction of lotus
fields. Using this method, the extraction accuracy of lotus was 96.3%. The Kappa coefficient was 0.926, which
is higher than those of the unsupervised K-means classification, Mahalanobis distance, and support vector
machine supervised classification, and demonstrates the potential of this method for extracting and mapping lotus
fields by remote sensing. 相似文献
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Purification of high-quality RNA from different strawberry tissues is often affected by the presence of high levels of contamination
by polysaccharides and phenolic compounds. With the protocol detailed here we describe for the first time total RNA purification
from petiole tissue. Treating the plants used as source of material with short-day light regime prior the extraction we are
able to obtain RNA suitable for further applications such as in vitro translation, RT-PCR, and RNA blot analysis. The yield
of total RNA extraction is significantly enhanced when tissue from plants grown under short-day photoperiodic condition is
used compared with that taken from plants grown under long day photoperiod. 相似文献