卷象科昆虫无损形态的DNA提取方法

提取DNA是所有分子生物学实验的第一步,但在昆虫分子生物学研究中,常规提取总DNA的方法不能保留昆虫所有的形态特征。显然,这不适用于对于体形较小的珍稀标本。以鞘翅目卷象科(Coleoptera:Attelabidae)昆虫为实验材料,提供了一种新的取材方法,取下整个腹部,消化其肌肉组织后收回骨化腹板。该方法可在提取昆虫总DNA的同时保存全部外形特征及其生殖器。     

硬蜱的采集与饲养

硬蜱为吸血节肢动物,是传播人和动物多种疾病的媒介。开展蜱传播疾病防治工作、硬蜱的生物学特性与分子生物学等方面的研究都需要硬蜱作实验材料。对硬蜱的人工采集方法、实验室活体饲养保存及固定标本保存技术进行了比较系统的阐述。     

桔小实蝇分子生物学研究中样品采集及保存处理的改进方法

介绍了一种果实蝇,桔小实蝇Bactroceradorsalis的虫样采集和保存处理的方法。利用一种简易的诱捕装置对于野外短时间、大面积采集实蝇虫样效果显著;同时,用TE溶液浸泡的干燥虫样适用于桔小实蝇的DNA抽提和PCR扩增。这种虫样采集和保存方法也适用于果实蝇属其它几类实蝇的分子生物学研究。     

鲜叶保存方法对茶树基因组DNA提取效果的影响

探讨一种简便有效的鲜叶保存方法,对克服茶树DNA研究中远距离采样的困难具有重要意义．为此,以槠叶齐品种的鲜梢为材料,分别采用-20℃、-70℃、硅胶脱水干燥3种方法保存样品,采用改进的CTAB法与SDS法分别对3种方法保存的样品及对照样(鲜叶)进行基因组DNA的提取与纯化,对所得DNA的质量进行多重检测的结果表明,硅胶脱水干燥法保存的样品与其他几种方法保存的样品一样,都提取获得了高质量的DNA,因此,硅胶脱水干燥法可作为远距离采样制备茶树DNA的一种较好的鲜叶保存方法,具有操作简单、经济实用、不受时空等条件限制的特点,该方法同样适用于其他植物．     

哺乳动物卵母细胞和胚胎的低温保存

超排技术的应用,可以得到几倍于正常排卵的卵母细胞或胚胎,为体外卵母细胞或胚胎的操作提供了丰富的材料。淘汰母畜或屠宰母畜的卵巢更是卵母细胞的一个重要研究资源。然而,如果供大于求,则需要一种较为满意的短期保存过剩卵母细胞或胚胎的方法。超低温(-196℃)冷冻固然是长期保存胚胎的良好方法,但对于只需短期(几个小时到几天)保存的胚胎而言,冷冻过程是烦琐的,且设备昂     

土壤真菌多样性的分子生物学研究方法

随着分子生物学技术的迅猛发展,越来越多的领域开始采用分子生物学研究方法。土壤真菌作为环境中不可或缺的要素,在生态环境中发挥着重要的作用。避开传统的分离培养方法,运用分子生物学方法对其研究具有深远的意义。本文主要介绍几种常见的土壤真菌分子生物学研究方法,从而为土壤真菌的研究提供理论依据。     

植物解剖材料的几种保存方法初探

本文介绍了植物解剖材料的三种保存方法:药液保存、直接冷冻保存、干燥保存。前一种方法适于野外植物采集,但所使用药品对环境和师生健康不利;后两种方法适于近距离或校园内植物采集,并且无毒无害。     

一种高效快速的鱼类标本基因组DNA提取方法

以95%酒精保存的黄鳝(M onopterus albus)和斑鳢(Channa maculates)标本为材料,采用先沉降DNA再去除杂质的方法从鱼类标本中提取基因组DNA。基因组DNA的琼脂糖凝胶电泳和紫外分光光度法检测以及PCR扩增结果显示,本方法提取的鱼类基因组DNA的电泳主带清晰明亮;A260/A280值在1.7830-2.0144之间;PCR扩增产物条带清晰明亮,且单一整齐没有拖带,表明本方法可从酒精保存的鱼类标本中提取比较纯净的DNA,能够满足一般分子生物学试验需要。与传统苯酚/氯仿法相比,本方法操作简单快速,避免了苯酚等物质对后续实验的影响,可作为一种常规动物组织DNA提取方法。     

农户保存与种质库保存的同近名地方稻品种的遗传多样性研究

通过对湖南农户自留种原地保存(简称农户保存)与异地低温保存在湖南种质库(简称种质库保存)的92份同近名水稻材料(7组,同近名材料6组)间的表现型及基因型进行分析,鉴定同近名品种间遗传距离,为之后有效保存、针对性供种、高效利用提供理论依据。结果表明,供试同近名材料间,及农户保存与种质库保存的同近名材料间在表现型性状上都有极显著差异。通过SSR标记基因型比较农户保存与种质库保存湖南同近名材料,发现除E组外,其他组都是农户保存的等位基因变异数小于种质库保存的等位基因变异数,说明农户保存的材料在年复一年的种植、收种过程中经历了自然选择和人工选择留种,进行了加代纯化。除C、E组外,在同近名组内SSR标记遗传相似系数显示,农户保存的材料间种质库保存的材料间农户保存材料与种质库保存材料,表明农户自留种保存的同近名水稻资源值得收集评价。     

不同方法保存的蜜蜂基因组DNA提取的比较

目的:找出适合DNA提取的昆虫标本保存方法。方法:用几种常用的昆虫标本保存方法对蜜蜂处理不同时间后,用蛋白酶K法对其基因组DNA进行提取和纯化,然后对提取产物做琼脂糖凝胶电泳及紫外吸收分析。结果:75%乙醇及冻存处理材料的基因组DNA得率较高,为7.13～8.85μg/g,电泳条带较亮;甲醛处理材料的基因组DNA得率较低,为1.50～3.21μg/g。结论:用75%乙醇及冻存处理蜜蜂较适合于其基因组DNA的提取,不宜用甲醛。     

Small-scale systems for in vivo drug delivery

Recent developments in the application of micro- and nanosystems for drug administration include a diverse range of new materials and methods. New approaches include the on-demand activation of molecular interactions, novel diffusion-controlled delivery devices, nanostructured 'smart' surfaces and materials, and prospects for coupling drug delivery to sensors and implants. Micro- and nanotechnologies are enabling the design of novel methods such as radio-frequency addressing of individual molecules or the suppression of immune response to a release device. Current challenges include the need to balance the small scale of the devices with the quantities of drugs that are clinically necessary, the requirement for more stable sensor platforms, and the development of methods to evaluate these new materials and devices for safety and efficacy.     

Computational discovery of nanoporous materials for energy- and environment-related applications

ABSTRACT

Our society is currently facing critical energy and environment issues, due to the consistent increase in the usage of fossil fuels and anthropogenic activities. One of the viable solutions is to develop better materials to enable more energy-efficient processes for various applications, including gas separations, energy storage, desalination, etc. Nanoporous materials such as zeolites, metal–organic frameworks (MOFs), and nanoporous graphene have drawn considerable attention as promising candidates in these applications owing to many of their favourable properties. Moreover, the tunability of porous materials results in essentially infinitely large number of possible candidates. While such vast materials space provides great opportunities, it also imposes a significant challenge on the selection of promising candidates. To this end, computational methods, such as molecular simulations, can play an important role in facilitating the discovery and design of optimal materials. In this review, we introduce several computational studies conducted for large-scale materials screenings for gas separations and for the discovery of novel membranes for water filtrations. Furthermore, in light of the importance of molecular force fields for reliable computational predictions, we also discuss some recent developments in this field. Overall, this article discusses the recent advances of computational material discoveries and methodology developments.     

论中药分子鉴定的方法和原则

中药的准确鉴定涉及到人民生命安全和切身利益。传统的鉴定方法,即感官评价、显微鉴定和理化鉴定均存在不同程度的局限性,而分子鉴定则为中药的快速和准确鉴定带来了新的契机。为了使中药的分子鉴定得到更加广泛和有效的应用,应该高度重视相关标准、规范的制订。本文提出了中药分子鉴定的一些主要方法：1)真伪品鉴定：特异聚合酶链式反应法;2)正品和替代品鉴定：DNA条形码鉴定法;3)多基源鉴定：群体遗传学分析法;4)产地鉴别：分子谱系地理学分析法。经上述方法仍无法鉴别的贵重药材可进一步应用人类亲子鉴定的方法,开发特异微卫星标记进行进一步的鉴定。     

On the Methods and Principles of MolecularIdentification of Chinese Herbs

Accurate identification of herbal medicinal materials is relevant to the safety of human life and economic interests. Traditional identification methods, including sensory evaluation, microscopic identification, physical and chemical identification, all have their limitations. Molecular identification brings a new opportunity for accurate identification of herbal medicinal materials. However, prior to its wide adoption, the methods and principles of molecular identification should be fully discussed. In this paper, we proposed a set of new methods for molecular authentication of herbal medicinal materials: 1) Identification between authenticity and adulteration by using specific polymerase chain reaction; 2) Identification between official herb and substitute by using the method of DNA barcoding; 3) Identification among multiple species of one official herbs by constructing genealogy among closely related species based on population genetics; 4) Identification among herbs of different geographical origins by phylogeography based analysis. For those that can not be identified by above four methods, more rapidly evolved markers such as microsatellite should be employed and individual based analysis could be adopted.     

玉米花药发育的细胞生物学与分子遗传学的研究方法

雄性不育技术在玉米杂种优势利用和杂交种生产中发挥着重要作用,玉米花药发育和雄性不育的细胞生物学与分子遗传学研究是雄性不育技术利用的前提和基础。玉米花药发育是一个复杂的生物学过程,需要孢子体基因与配子体基因的协同表达调控。从玉米花药的形态结构、花药发育时期的划分、花药败育类型、花药发育的细胞学研究方法、花药发育的组学研究方法、花药发育分子遗传学研究方法等方面进行综述,以期为玉米核不育机制研究与雄性不育技术产业化应用提供方法学指导。     

A comparison of different methods for preserving plant molecular materials and the effect of degraded DNA on ddRAD sequencing

Obtaining high-quality plant materials for experiments is challenging for many research projects. Therefore, it is of special importance to determine the best method for preserving biological macromolecules like DNA, which degrade over time. Although some research has demonstrated that DNA degradation has little effect on traditional molecular markers, the effects of DNA degradation on ddRADseq, a popular reduced-representation sequencing technology, have not been adequately investigated. In this study, we first chose six woody bamboo species (Bambusoideae, Poaceae) to explore appropriate methods for preserving molecular materials with two DNA extraction approaches. Then we sequenced twenty-one bamboos and examined the effects of DNA quality on data generation using the ddRAD-seq technique (MiddRAD-seq). Finally, we reconstructed phylogenies of twenty woody bamboo species. We found that the integrity of dry-powdered DNA was preserved longer than that of TE-dissolved DNA, regardless of whether the DNA was extracted by a modified CTAB protocol or DNAsecure plant kit. The ddRAD-seq data were robust, except when DNA was severely degraded. In addition, we resolved the phylogenetic positions of the sampled Phyllostachys spp. Our results suggest that dry-powdered DNA is the most appropriate preservation method for plant molecular materials. Furthermore, a moderate level of DNA degradation has little effect on reduced representation sequencing techniques represented by ddRAD-seq.     

A protocol for high-throughput extraction of DNA from rice leaves

Current DNA isolation methods have limitations between speed and purity in high-throughput molecular genetic analysis such as gene mapping and marker-assisted selection programs. We have optimized a simple and rapid method for isolating high-quality genomic DNA from rice that significantly minimizes time and the use of laboratory materials. One person can process as many as 384 samples in 2 h. The isolated DNA is suitable for polymerase chain reaction-based techniques and is stable for no less than 6 mo of storage at 4°C.     

Synthesis of hierarchical materials

A major goal of material science is to produce hierarchical materials that are ordered on all length scales, from the molecular (1-100 A) via the nano (10-100 nm) to the meso (1-100 microm). In these materials, the larger-scale properties can be controlled by choosing molecular characteristics. Methods developed to produce three-dimensional, bulk-like hierarchical structures include biomimetic methods, which use polypeptides as building blocks, and amphiphile and colloidal templating, which use amphiphilic or colloidal mesophases as templates for inorganic mesoporous materials. Designing finite mesostructures with a given geometry still remains a challenge.