CTAB法提取中草药材滇紫草细胞的总RNA

介绍一种适合富含次生产物的中草药材滇紫草细胞总RNA的提取方法——CTAB法。采用CTAB作为去污剂,分别用氨仿和水饱和酚反复抽提以及LiCl沉淀以去除蛋白质、碳水化合物和次生代谢物等杂质,最后用纯乙醇沉淀获得RNA。该方法不但能获得完整性好和纯度高的总RNA,而且操作简单、快速、成本低廉,对其它富含次生物质的植物特别是中草药材组织总RNA的提取具有借鉴意义。     

自然干燥紫萼藓总RNA提取方法

介绍一种适合富含酚类、萜类等次生物质的干燥紫萼藓的总RNA的提取方法—SDS/酸酚法。采用SDS做为去污剂,用水饱和酚、氯仿和异戊醇进行抽提以去除蛋白、酚类等次生物质,醋酸钾和无水乙醇去除多糖等物质,最后LiCl沉淀获得总RNA。该方法不但获得了完整性好和纯度高的RNA,而且操作简单,成本也较低,对其他富含酚类、萜类等次生物质的干燥植物组织的总RNA的提取具有借鉴意义。     

淡水育珠蚌外套膜提取总RNA的改良方法

通过对Trizol法加以改进,提取淡水珍珠蚌外套膜组织中的总RNA。经预处理后,在异丙醇沉淀RNA时加入高浓度的盐溶液,用75%的酒精2次洗涤RNA。用紫外分光光度法和1%琼脂糖凝胶电泳鉴定所提取的RNA。结果表明,改良法获得的总RNA完整、纯度高,改良Trizol法是一种从淡水育珠蚌外套膜组织中提取总RNA的高效、便捷、可靠的方法。     

一种从富含多糖的玉米幼穗中提取RNA的方法

介绍了一种大量提取玉米(Zeamays L.)幼穗总RNA的有效方法。由于玉米幼穗富含多糖,用普通的RNA提取方法和Trizol很难获得高质量的RNA。在热酚法提取RNA的基础上,通过在提取缓冲液中加入低pH值的醋酸钾来去除多糖。所得到的RNA获得率高,质量好,条带完整,实验证明可直接用于RT-PCR、微阵列等各项后续分子生物学实验。     

一种从富含多糖的玉米幼穗中提取RNA的方法

介绍了一种大量提取玉米(Zea mays L.)幼穗总RNA的有效方法。由于玉米幼穗富含多糖, 用普通的RNA提取方法和Trizol很难获得高质量的RNA。在热酚法提取RNA的基础上, 通过在提取缓冲液中加入低pH值的醋酸钾来去除多糖。所得到 的RNA获得率高, 质量好, 条带完整, 实验证明可直接用于RT-PCR、微阵列等各项后续分子生物学实验。     

一种高效经济的高质量植物RNA提取方法

建立了一种高效经济的植物RNA提取方法.在提取缓冲液中加入蔗糖、氯化钾和镁离子以提供对RNA分子的保护.破碎后的细胞于提取缓冲液中裂解后,用酚/氯仿变性并去除内源RNA酶和其他蛋白质,而后用pH 5.6 的NaAc沉淀RNA.用该方法提取RNA的得率较高,经电泳检测,RNA的完整性很好.RNA印迹分析和RT-PCR也都得到很好的结果.该方法还使实验成本大大降低.     

一种适用范围广的总RNA提取方法

介绍一种RNA提取方法,该方法以SDS、氯仿和Tris苯酚为主要提取试剂,以LiCl和乙醇为RNA沉淀试剂。分别以柽柳(木本植物)、星星草(草本植物)、天牛（昆虫）、酿酒酵母和白腐菌（真菌）为RNA提取材料,用该方法成功地提取出了它们的总RNA。获得的RNA条带清晰,A₂₆₀/A₂₈₀ 在1.8以上。通过对LiCl和乙醇沉淀RNA的效果分析表明,该方法可在10 min内完全沉淀RNA,同时也可以同时获得纯度较高的DNA。提取的RNA质量可满足cDNA文库构建,基因芯片探针标定和RT-PCR等对RNA质量要求较高的分子生物学操作,说明这是一种应用范围广的RNA提取方法。     

仿刺参体壁总RNA提取方法的建立

目的:通过对TRIzol一步法进行改进,建立一种从富含胶原蛋白、多糖及色素的仿刺参体壁提取总RNA的有效方法。方法:样品在液氮中研磨并用TRIzol匀浆后再进行抽提;对TRIzol一步法提取的总RNA进行DNaseⅠ消化和酚氯仿抽提,用2.5mol/L的醋酸钾沉淀,并加入适量糖原(10mg/mL)与RNA共沉淀。结果:琼脂糖凝胶电泳和紫外分光光度法以及RT-PCR检测结果表明,改进的方法能够有效去除基因组DNA、蛋白、多糖及色素的污染,RNA的产率提高。结论:制备的总RNA纯度高,完整性好,能够满足mRNA差异显示RT-PCR等分子生物学研究的要求,是一种提取仿刺参体壁及其他富含黏多糖、胶原蛋白和色素的动物组织总RNA的有效方法。     

毛尖紫萼藓总RNA的提取方法研究

介绍一种提取苔藓植物毛尖紫萼藓(Grimmia pilifera P.Beauv)总RNA的方法。以新鲜的紫萼藓为材料,采用改良的SDS的方法,以氯化锂为沉淀试剂,成功地提取了该植物的总RNA。并与其他方法作了对比,结果发现该方法获得的RNA条带清晰、完整性好、纯度高、DNA污染小。可满足反转录及RT-PCR等实验要求。     

大青杨叶片总RNA的快速提取方法

目的:寻求快速提取大青杨叶片总RNA的方法。方法:分别用改良CTAB法、改良SDS法、改良TRIzol法及某公司总RNA提取试剂盒提取大青杨叶片总RNA,并用紫外光谱分析、凝胶电泳方法对提取的总RNA进行鉴定。结果:用改良CTAB法和改良TRIzol法能有效地去除蛋白及多糖,提取到的总RNA纯度高,D260nm/D280nm分别为2.05和1.78,RNA的完整程度优于试剂盒法。结论:改良CTAB法为大青杨叶片总RNA的最佳提取方法。     

用改良CsCl密度梯度离心法提取石蒜叶片和鳞茎总RNA

石蒜[Lycoris radiata（L’Her．）Herb．]是重要的药用植物,其次生代谢产物加兰他敏（galantamine）在临床上的应用非常广泛,是治疗阿尔茨海默氏病的首选药物之一。尽管有关加兰他敏化学合成途径已有许多报道,但由于纯化学合成的成本太高,目前仅限于实验室合成,尚不能用于工业化生产。提高石蒜本身的加兰他敏含量是获得加兰他敏最为经济和环保的方法。因此,研究加兰他敏生物合成的分子机制对于提高其含量具有重要意义。     

Improved technique for isolating RNA from tobacco tissues

We have developed a much-improved method for isolating RNA from tobacco tissue. The novel component of the described RNA isolation method is the addition of lithium chloride to the extraction buffer. Following that, the RNA was homogenized with phenol/chloroform and precipitated in ethanol. This isolation technique provided highly reproducible and good quality RNA within 2 h.     

提取航天诱变向日葵种子总RNA的一种有效方法

目的:从航天诱变向日葵种子中提取高质量的总RNA.方法:采用改进的SDS法,提取缓冲液与氯仿同时作用液氮研磨材料后,用酸酚-氯仿抽提一次,经LiCl过夜沉淀、DNase I处理、1/2体积的无水乙醇沉淀多糖,最后加入1/10体积的醋酸钠和2倍体积的无水乙醇沉淀总RNA,用琼脂糖凝胶电泳与紫外分光光度法测定产量与纯度,用...     

从血液中提取总RNA的一种快速高效方法

血液中含有大量的RNA酶 ,可引起RNA的降解 .防止RNA酶的降解 ,是保证所得RNA片段完整的关键 .目前提取RNA的方法较多 ,但有些方法尚不能完全防止RNA降解 .将TRIZOL方法稍加改进 ,将TRIZOL与异硫氰酸胍联用提取血液淋巴细胞总RNA .琼脂糖凝胶电泳结果表明 ,其 2 8SRNA与 18SRNA的比值为 2∶1,优于单独使用其中任何一种试剂者 .此方法同样适用于从其它细胞中提取RNA .     

Extraction of DNA,RNA, and glycogen from oysters

The macromolecules DNA, RNA, and glycogen (CHO)_N were extracted with phenol from eggs, sperm, and the combined gill, mantle, and digestive gland tissues of oysters, using methods effective for the HeLa cells. From the eggs, most of the (CHO)_N and RNA coprecipitated in 20% ethanol whereas the DNA precipitated from 50% ethanol solutions. From the combined tissues all three macromolecules precipitated in 20% ethanol whereas from sperm they precipitated from 50% ethanol solutions. The DNA preparation from sperm was viscous and white but contained mostly (CHO)_N and some RNA. However, the DNA-(CHO)_N-RNA preparation was rendered insoluble in saline by CaPO₄ treatment and was not toxic for mammalian kidney cells in tissue culture. Thus, the methods do not yield pure nucleic acids but appear suitable for attempts to extract infectious DNA from oysters.     

Selective precipitation of ribonucleic acid from a mixture of total cellular nucleic acids extracted from cultured mammalian cells

A simple and reproducible method is described for precipitating RNA selectively from total mammalian-cell nucleic acids extracted by the phenol-sodium dodecyl sulphate procedure at pH8.0. Under specified conditions bulk RNA is precipitated almost quantitatively whereas bulk DNA remains in solution. Minor components of RNA (detected by pulse-labelling and chromatography on methylated albumin-kieselguhr) and rapidly labelled components of DNA containing single-stranded regions are also precipitated. The usefulness of the method is discussed in the context of isolating separately both RNA and DNA from cultured cells that are difficult to obtain in quantity.     

介绍一种简单高效的植物总RNA提取方法

在液氮中研磨小麦幼叶和不同发育时期的种子,经含0.1% SDS和0.1%十二烷基肌氨酸钠(LDS)的尿素缓冲液裂解后,醋酸钠和氯仿沉淀变性蛋白质,异丙醇沉淀核酸,溶解后经2.5mol/L LiCl沉淀总RNA,洗涤后就可得到高质量的总RNA,其OD260/OD280 为2.05~2.10,28S和18S RNA带清晰,叶片总RNA还可得到23S和16S RNA带,产率可达5mg RNA/10g材料。当使用含1% SDS和1% LDS的尿素缓冲液裂解材料时,则可用于DNA的分离提取,其分子大小可达50~100kb以上。 Abstract:Wheat leaf and seeds at different development stages had been squashed in liquid nitrogen,then lysised by urea buffer which contains 0.1% SDS and 0.1% LDS,denatured protein had been removed by NaAc and chloroform precipitation,total RNA was further purified by LiCl.The RNA we obtained had sharp bands of 28S and 18S after agarose gel electrophoresis,23S and 16S RNA bands can also be seen clearly in leaf RNA extract,the value of OD260/OD280 of RNA was 205~210.5mg RNA can been isolated from 10g leaf of wheat.This method can also been used in high molecular weight DNA isolation but the concentration of SDS and LDS must be increased to 1%.     

Using a commercial DNA extraction kit to obtain RNA for RT-PCR from starchy rice endosperm

The extraction of RNA from a starchy plant material, such as many common food grains, is difficult, and especially so from the mature endosperm of rice. Most commercial RNA kits are not suitable for starchy materials. Traditional RNA extraction procedures, in addition to being laborious and time consuming, leave hazardous organic wastes that result in expensive disposal costs. Interestingly, the numerous commercial DNA isolation kits now available often include directions for eliminating co-isolated RNA. This indicated an approach to obtain the generally unwanted RNA by-product by treating the total extraction product to intentionally retain RNA. A method was developed by which a two-step DNase procedure was applied to the product of the Cartagen Food DNA extraction kit that eliminated the DNA but left the co-extracted RNA. This modified procedure was compared with several other commercial and standard methods that are promoted as being able to work under high polysaccharide conditions. Successful extraction was determined by the production and amplification of cDNA by RT-PCR of actin. Extraction was successful from milled rice, as well as from cornmeal and wheat flour. The modification provides an RNA extraction method that is quick, easy, and inexpensive, and also eliminates the production of hazardous wastes.