A Method for Isolation of Chloroplast DNA and Mitochondrial DNA from Sunflower

We present a method for isolation of chloroplast and mitochondrial DNA from sunflower seedlings. The protocol includes: organelle isolation, deoxyribonuclease treatment, lysis, deproteinisation and a final DNA purification with sodium dodecyl sulphate and potassium acetate. The organelle DNA yield is 5–10 micrograms per gram of tissue and the DNA is fully restrictable. The technique is inexpensive and appropriate for the isolation of multiple samples of organelle DNA from a small amount of tissue.     

水稻基因组DNA微量提取

随着植物学向分子水平的深入发展,研究中经常需要获得高质量的植物DNA样品,因此,建立植物DNA提取与纯化的常规实验方法对教学和科研都显得非常必要。介绍一种快速提取微量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,可满足以PCR扩增为基础的实验需要。     

High throughput isolation of DNA and RNA in 96-well format using a paint shaker

Most steps in plant nucleic acid isolation are easily adapted to 96-well format; however, tissue disruption typically is performed on samples individually and often is the rate-limiting step in sample processing. We have found that DNA and RNA isolation from Arabidopsis tissue can be carried out in 96-well format using a paint shaker fitted with an adapter for tissue disruption.     

A rapid and inexpensive method for isolation of total DNA from dehydrated plant tissue

We describe an inexpensive method for dehydration of plant tissue and extraction of high molecular weight DNA. Tissue is dried for 12 to 24 hours in a food dehydrator and subsequently powdered for DNA extraction. Dicot tissue can be powdered in centrifuge tubesen masse using a commercial paint mixer and glass beads. With the use of the paint mixer, tissue never touches common surfaces that might lead to cross contamination, a potential benefit when the DNA is to be used for PCR reactions. The DNA is of a quality equal to that obtained from either lyophilized or fresh frozen tissue (commonly used in many labs). The advantages of the described procedure are that it is fast, does not require expensive equipment (e.g., lyophilizer) and can be used in situations where large numbers of samples must be extracted.     

一种高效提取猕猴桃DNA和RNA的方法

在总核酸提取方法(PS法)的基础上,经多次实践改进,得出一种以高盐低pH的HAc-NaAc缓冲体系提取总核酸的简便方法,可以从富含多糖、多酚时猕猴桃叶片和花蕾中提取同时含有DNA和RNA的总核酸.所得的总核酸在LiCl溶液中选择性沉淀RNA,从而有效地分离出DNA和RNA样品.紫外分光光度法和琼脂糖凝胶电泳分析表明,所提取的DNA和RNA具有较高的纯度和完整性.通过样品DNA的PCR和样品RNA的RT-PCR,认为所提取的DNA样品和RNA样品能够满足分子生物学试验的基本要求.     

小型昆虫DNA提取时匀浆方法的改进

应用改进的方法方便地提取了高质量的单头蚜虫基因组DNA ,有助于解决小型昆虫研究工作中单头虫体的DNA提取困难问题     

A modified DNA isolation protocol for obtaining pure RT-PCR grade RNA

We provide a simple but very efficient method for RNA preparation from Saccharomyces cerevisiae based on a standard chromosomal DNA isolation protocol. The method yields DNA-free total RNA, including mRNA, rRNA, and tRNA but can easily be adjusted to considerably enrich low molecular weight RNAs, such as tRNAs and the small rRNA species (5S and 5.8S). The procedure was proven and validated by verification of cDNAs belonging to four different genes, two of which encoding polypeptides and two tRNA genes. Besides its simpleness, the method is further advantagous in terms of safety (omitting hazardous phenol) and cost efficiency.     

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。     

Isolation of High Quality DNA and RNA from Leaves of the Carnivorous Plant Drosera rotundifolia

Drosera rotundifolia belongs to the family of the sundews, a large group of carnivorous plants that carry stalked glands on the upper leaf surface to attract, trap and digest insects for food. Therefore, such plants can live in relatively poor ecosystems. They are frequently used as medicinal herbs and have various other interesting characteristics associated with them. In attempts to evaluate the gene pool of these plants, we experienced that many published protocols for nucleic acid isolation failed to yield DNA and RNA of sufficient quality for analysis. Therefore, we have developed CTAB (hexadecyltrimethylammoniumbromide)-based extraction protocols for the routine isolation of high-quality DNA and RNA from small amounts of in vitro-grown Drosera rotundifolia leaves. The methods developed are simple, fast and effective. The obtained DNA could be analyzed by PCR, restriction endonucleases and DNA gel blotting, and the obtained RNA was of sufficient quality for RT-PCR and RNA gel blotting.     

线粒体DNA和疾病

人线粒体DNA是含有16569 bp的闭环双链分子.它为13种氧化磷酸作用酶的亚单位、结构rRNAs和tRNAs编码.近年来很多引起人类疾病的线粒体DNA突变已被确定,如眼盲、耳聋、心力衰竭和人类退行性疾病等.线粒体DNA疾病可能比先前想象的多.     

Rapid small-scale DNA isolation from filamentous cyanobacteria

Abstract A rapid small-scale DNA isolation procedure is described for the filamentous cyanobacteria, which yields enough chromosomal and plasmid DNA for restriction endonuclease digestions, Souther hybridizations, and electroelution from gels for further manipulation. DNA from seven strains of cyanobacteria were isolated and analyzed on agarose gels.     

Isolation of functional RNA from small amounts of different grape and apple tissues

An efficient, simple, and small-scale procedure for isolating functional ribonucleic acid (RNA) was successfully applied to many different tissues of grape and apple. These woody plants are rich in polyphenolic compounds and polysaccharides that could impair the RNA extraction. The method chosen is based on the use of hot borate buffer at alkaline pH supplemented with several adjuvants and followed by selective precipitations. Starting with only 0.4 g of fresh tissue and working with small tubes (2 mL), we were able to obtain good yields of high-quality RNA suitable for further applications. The procedure can be proposed for many applications, and it is particularly highly recommended when isolating RNA from a large number of samples.     

线粒体DNA突变与相关人类疾病

在过去的20年里, 人们发现线粒体DNA(mitochondrial DNA, mtDNA)突变与多种人类疾病相关, 其致病范围从单器官组织损害到多系统受累。文章目的在于探讨mtDNA突变与人类疾病的关系。文章重点论述: (1)线粒体遗传学特征; (2) mtDNA突变与人类遗传性疾病; (3)体细胞mtDNA突变在衰老和肿瘤中的作用; (4)mtDNA疾病的诊断和治疗。     

人线粒体RNA加工及调控

线粒体是细胞内氧化磷酸化（oxidative phosphorylation,OXPHOS）和合成三磷酸腺苷（adenosine triphosphate,ATP）的细胞器，是细胞能量代谢的“动力工厂”。线粒体几乎存在于所有真核生物中，参与细胞凋亡、钙稳态以及先天免疫反应的调节等过程，对细胞行使正常的生理功能至关重要。线粒体是半自主细胞器，拥有自身的基因组DNA，编码37个基因，包括2个rRNA基因、13个m RNA基因和22个tRNA基因。线粒体的基因表达需要经过复杂的转录和转录后加工过程，包括多顺反子RNA的切割、RNA的修饰以及RNA的末端加工等过程。异常的线粒体RNA加工会导致线粒体RNA表达谱发生变化、线粒体翻译紊乱、线粒体功能失常等，从而造成多种线粒体相关疾病。本文综述了线粒体DNA的转录、RNA转录后加工以及影响RNA加工的因素方面的最新研究进展。     

Conservation of ribosomal RNA gene arrangement in the mitochondrial DNA of angiosperms

In six different angiosperms, mitochondrial genes for 18S and 5S rRNAs were found to be closely linked but distant from mitochondrial 26S rRNA genes.This is paper no. 5 in the series Organization and Expression of the Mitochondrial Genome of Plants     

Isolation of Functional RNA from Periderm Tissue of Potato Tubers and Sweet Potato Storage Roots

A reliable and efficient protocol is given for the isolation of mRNA from the periderm of potato tubers and sweet potato storage roots. The method relies on a urea-based lysis buffer and lithium chloride to concentrate total RNA away from most of the cytoplasmic components and to prevent oxidation of phenolic complexes. To enhance the physical separation of the RNA from other macromolecular components, the RNA fraction was incubated in the presence of the cationic surfactant Catrimox-14. Poly(A)⁺ mRNA was separated from total RNA and other contaminants by using Promega's MagneSphere technology. The mRNA was suitable for cDNA library construction and RNA fingerprinting.