大鼠第三脑室室管膜NADPH-d阳性伸展细胞的形态与分布

应用 Ｎ Ａ Ｄ Ｐ Ｈｄ 组织化学方法研究了大鼠第三脑室视前区室管膜的伸展细胞⒚结果表明：１）第三脑室视前区室管膜存在 Ｎ Ａ Ｄ Ｐ Ｈｄ 阳性的伸展细胞,其基突伸向视前区并与神经元或毛细血管相接触;２） Ｎ Ａ Ｄ Ｐ Ｈｄ 阳性的伸展细胞在第三脑室侧壁常见、室底少见、室顶未发现其存在;３） Ｎ Ａ Ｄ Ｐ Ｈｄ 阳性的伸展细胞的形态与分布,在雌、雄大鼠间不存在明显的性别差异⒚尽管 Ｎ Ａ Ｄ Ｐ Ｈｄ 阳性的伸展细胞的生理功能不十分清楚,但本研究为伸展细胞作为脑脑脊液环路的一部分,介导下丘脑对脑脊液中化学变化的感受提供了形态学依据,并提示一氧化氮可能参与了这一过程⒚     

花生种子高纯度DNA的提取（简报）

花生种子高纯度ＤＮＡ的提取（简报）王艳,何军贤,傅家瑞（中山大学生命科学学院,广州５０２７５）关键词花生;种子;ＤＮＡ;十六烷基三甲基溴化铵ＲＡＰＩＤＡＮＤＥＦＦＩＣＩＥＮＴＰＵＲＩＦＩＣＡＴＩＯＮＯＦＤＮＡＦＲＯＭＰＥＡＮＵＴ（ＡＲＡＣＨＩＳＨＹＰ...     

眼镜蛇毒腺cDNA文库的构建

目的　蛇毒含有多种生物活性成分, 从天然蛇毒中提取分离蛇毒有效成分受到蛇毒资源和质量的限制。为开发蛇毒有效成分的基因工程产品, 本研究构建了蛇毒腺的ｃ Ｄ Ｎ Ａ 文库, 为进一步筛选、克隆和表达蛇毒有关基因做准备。　方法　从眼镜蛇（ Ｎａｊａ ｎａｊａ ａｔｒａ ）毒腺中提取ｍ Ｒ Ｎ Ａ,经反转录合成ｃ Ｄ Ｎ Ａ后, 以λｇｔ１０ 噬菌体为载体, 构建非表达的ｃ Ｄ Ｎ Ａ 文库。　结果　蛇毒腺ｃ Ｄ Ｎ Ａ 非表达文库库容量为２×１０６ｐｆｕ／μｇ 重组子, 经大肠杆菌 Ｃ６００ ｈｌｆ 菌株平皿测定, 重组率为 ７０％ 。　结论　经平板鉴定和 Ｐ Ｃ Ｒ 快速鉴定表明, 所构 ｃ Ｄ Ｎ Ａ 文库达到建库要求, 能够用于目的基因筛选和克隆表达。     

丹顶鹤与白枕鹤的领域比较研究

丹顶鹤与白枕鹤的领域比较研究ＡＣＯＭＰＡＲＡＴＩＶＥＳＴＵＤＹＯＮＴＥＲＲＩＴＯＲＩＥＳＯＦＷＨＩＴＥＮＡＰＥＤＣＲＡＮＥＡＮＤＲＥＤＣＯＲＯＷＮＥＤＣＲＡＮＥ关键词丹顶鹤白枕鹤领域ＫｅｙｗｏｒｄｓＲｅｄｃｒｏｗｎｅｄｃｒａｎｅ,Ｗｈｉｔｅｎ...     

吉林梅花鹿体尺,体重聚类与主成分分析

吉林梅花鹿体尺、体重聚类与主成分分析ＤＬＵＳＴＥＲＡＮＤＰＲＩＮＣＩＰＡＬＣＯＭＰＯＮＥＮＴＡＮＡＬＹＳＩＳＯＦＴＨＥＣＨＡＲＡＣＴＥＲＩＳＴＩＣＳＡＮＤＷＥＩＧＨＴＯＦＪＩＬＩＮＳＩＫＡＤＥＥＲ￥ＬＩＨｅｐｉｐｇ;ＢＩＮＧＧｕｏｌｉａｎｇ;ＰＡＮＧ...     

被子植物核型胚乳细胞化机理研究的现状

被子植物核型胚乳细胞化机理研究的现状李师翁（庆阳师专生物系,甘肃西峰７４５０００）ＡＤＶＡＮＣＥＳＩＮＴＨＥＳＴＵＤＩＥＳＯＮＴＨＥＭＥＣＨＡＮＩＳＭＯＦＣＥＬＬＵＬＡＲＩＺＡＴＩＯＮＯＦＮＵＣＬＥＡＲＥＮＤＯＳＰＥＲＭＩＮＡＮＧＩＯＳＰＥＲＭＳＬｉ...     

东北鼢鼠种群生长指标的主分量分析

东北鼢鼠种群生长指标的主分量分析ＰＲＩＮＣＩＰＡＬＣＯＭＰＯＮＥＮＴＡＮＡＬＹＳＩＳＯＦＧＲＯＷＴＨＩＮＤＥＸＯＦＰＯＰＵＬＡＴＩＯＮＦＯＲＭＡＮＣＨＵＲＩＡＮＺＯＫＯＲ（ＭＹＯＰＡＬＡＸＰＳＩＬＵＲＡＳ）东北鼢鼠（Ｍｙｏｐａｌａｘｐｓｉｌｕｒａｓ...     

大鼠隔区一氧化氮合酶阳性神经元的分布和脑缺血后的变化

大鼠隔区一氧化氮合酶阳性神经元的分布和脑缺血后的变化ＤＩＳＴＲＩＢＵＴＩＯＮＡＮＤＩＳＣＨＥＭＩＡＩＮＤＵＣＥＤＣＨＡＮＧＥＳＯＦＮＩＴＲＩＣＯＸＩＤＥＳＹＮＴＨＡＳＥＰＯＳＩＴＩＶＥＮＥＵＲＯＮＳＩＮＴＨＥＳＥＰＴＡＬＡＲＥＡＯＦＲＡＴ关键词大鼠...     

在长江沙洲上搁浅的中华白海豚

在长江沙洲上搁浅的中华白海豚ＳＴＲＡＮＤＩＮＧＯＦＡＮＩＮＤＯ┐ＰＡＣＩＦＩＣＨＵＭＰ┐ＢＡＣＫＥＤＤＯＬＰ┐ＨＩＮＯＮＡＳＡＮＤＢＡＮＫＩＮＴＨＥＹＡＮＧＴＺＥＲＩＶＥＲ中华白海豚（Ｓｏｕｓａｃｈｉｎｅｎｓｉｓ,Ｏｓｂｅｃｋ）分布在西太平洋和印度...     

植物胚胎学实验方法（七）同时显示胚中贮藏的淀粉,蛋白质和脂类的永久制片法

植物胚胎学实验方法（七）同时显示胚中贮藏的淀粉、蛋白质和脂类的永久制片法胡适宜（北京大学生物系,北京１００８７１）ＭＥＴＨＯＤＯＦＰＲＥＰＡＲＡＴＩＯＮＯＦＳＬＩＤＥＳＦＯＲＳＩＭＵＬＡＮＥＯＵＳＤＥＭＯＮＳＴＲＡＴＩＯＮＳＯＦＳＴＡＲＣＨＧＲＡＩＮ...     

Chelex without boiling, a rapid and easy technique to obtain stable amplifiable DNA from small amounts of ethanol-stored spiders

DNA barcoding projects require high-throughput generation of sequence data to assemble the comprehensive reference databases that are required to perform large-scale biodiversity inventories and molecular ecology studies. With the advent of new sequencing technologies, the extraction step, which often requires a considerable amount of time and money, represents a significant bottleneck in many studies. Here, we present a one-step Chelex double-stranded DNA extraction protocol that is quick, cheap, easy and works with a small quantity of ethanol-stored tissue. We developed this protocol by removing the denaturation step appearing in classic methods. This modification reduces the number of handling steps to one, thus simplifying the extraction procedure and reducing the risk of sample contamination, and yields double-stranded DNA instead of the single-stranded form that classical Chelex extraction protocols usually release. DNA obtained through our method is then suitable for long-term conservation (over 1.5 years). We tested our protocol on a highly diverse genus of spiders comprised of mainly very small species. We also apply the method to two other genera of spiders, one with average size species, the other one with giant species, to test the efficacy of the method with varying amounts of input tissue. We also discuss the advantages and limitations of this DNA extraction technique when working with arthropods.     

转基因植物快速检测方法的研究

本试验对转基因植物检测中的DNA提取和PCR扩增程序作了改进。经试验,本研究建立的DNA快速提取法与目前广泛使用的CTAB法相比更为简便,快速和经济,提取的DNA质量主扩增效果无明显差异,可用于多种转基因植物,多种植物组织的DNA提取,利用复合PCR法可在同一反应管中同步检测35N,NOS及CP4－EPSPS基因,明显提高了检测效率。应用本试验建立的DNA快速提取－复合PCR扩增－银染检测技术可在6小时内得出结果,达到了快速,简便,灵敏,可靠的检测目的。     

A RAPID PCR-QUALITY DNA EXTRACTION METHOD IN FISH

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Testing compatibility between molecular and morphological techniques for arthropod systematics: a minimally destructive DNA extraction method that preserves morphological integrity, and the effect of lactic acid on DNA quality

Three practical aspects related to the preservation and destruction of DNA and/or morphological characters of spiders were examined: potential morphological damage during non-destructive DNA extraction was assessed by counting trichobothria, a fragile sensorial feature found on spider legs; the effect on yield of non-destructive DNA extraction; and whether possible DNA degradation is caused by residues of lactic acid, which is used as a temporary mounting medium for the study of morphological structures in spiders and insects. Destructive extractions yielded higher amounts of DNA than non-destructive methods. However, non-destructive methods yielded usable amounts of DNA while leaving delicate trichobothria intact. Of the non-destructive extractions, a longer digestion period (36 h vs. 12) yielded higher amounts of DNA and did not damage trichobothria. Lactic acid did not induce short-term DNA degradation or inhibit PCR reactions, even at high concentrations. These results show compatibility between molecular and morphological requirements without compromising DNA quality or specimen integrity.     

Purification of human genomic DNA from whole blood using sodium perchlorate in place of phenol

We have developed a new, rapid method for the extraction of human genomic DNA from whole blood samples. Traditionally, genomic DNA has been extracted from blood by overnight proteinase K digestion of lysed peripheral lymphocytes followed by phenol/chloroform extraction. In addition to being time consuming, the use of phenol involves inherent risks due to the toxic nature of the reagent. Our method for the extraction of DNA from whole blood uses sodium perchlorate and chloroform instead of phenol with a significant time savings realized as well as fewer hazards to the technician. Furthermore, DNA prepared by this new method is an excellent substrate for restriction endonuclease digestion and Southern hybridization analysis.     

一种快速、经济提取外周凝血基因组DNA方法的建立

目的：建立一种经济、快速且高质量提取人体外周凝血DNA的方法。方法：摸索最佳的匀浆条件,对外周凝血块进行匀浆,采用Ⅺ法对匀浆液进行基因组DNA的提取,通过凝胶电泳、单重PCR和多重PCR检测凝血基因组DNA的提取产量和质量。并分别与常规的凝血基因组DNA提取方法,即蛋白酶K消化法,以及提取抗凝血基因组DNA的Ⅺ法进行比较分析。结果：最佳的匀浆条件为：39000map,15秒。在此条件下提取的基因组DNA完整性好,纯度和产量与蛋白酶K消化法提取凝血DNA和KI法提取抗凝血DNA的结果相比,没有统计学差异。单重PCR和多重PCR也获得了理想的扩增结果。结论：与常规的外周凝血提取方法相比（蛋白酶K消化法）,本方法节省了时间和成本,能快速、经济、有效地提取外周凝血基因组DNA,可用于后续的科研和临床诊断需要,解决了部分科研机构血液基因组DNA的样本来源问题。     

一种适于转基因水稻PCR检测的微量DNA快速提取法

对已报道的小麦基因组DNA快速提取方法的部分步骤进行了简化,在水稻上进行了尝试。结果表明,简化法提取的水稻基因组DNA完整性好,PCR扩增效果与试剂盒提取法无明显的差异,结果稳定可靠;而且整个提取过程操作简单、花费时间少,样品用量少,仅需5-10mg,适用于大规模转基因水稻的PCR检测。     

一种快速的胚胎组织总RNA的提取方法

简要介绍一种改进的提取人体器官组织总RNA的方法,该方法具有费用低,快速简便,重复性好的优点,提取的RNA无DNA等污染物,完全能满足基因表达研究的需要。     

Rapid, one-step DNA extraction for insect pest identification by using DNA barcodes

Early detection of economically important insects is critical to preventing their establishment as serious pests. To accomplish this, tools for rapid and accurate species identification are needed. DNA barcoding, using short DNA sequences as species "genetic identification tags," has already shown large potential as a tool for rapid and accurate detection of economically important insects. DNA extraction is the critical first step in generating DNA barcodes and can be a rate-limiting step in very large barcoding studies. Consequently, a DNA extraction method that is rapid, easy to use, cost-effective, robust enough to cope with range of qualities and quantities of tissue, and can be adapted to robotic systems will provide the best method for high-throughput production of DNA barcodes. We tested the performance of a new commercial kit (prepGEM), which uses a novel, streamlined approach to DNA extraction, and we compared it with two other commercial kits (ChargeSwitch and Aquapure), which differ in their method of DNA extraction. We compared performance of these kits by measuring percentage of polymerase chain reaction (PCR) success and mean PCR product yield across a variety of arthropod taxa, whichincluded freshly collected, ethanol-preserved, and dried specimens of different ages. ChargeSwitch and prepGEM performed equally well, but they outperformed Aquapure. prepGEM was much faster, easier to use, and cheaper than ChargeSwitch, but ChargeSwitch performed slightly better for older (> 5-yr-old) dried insect specimens. Overall, prepGEM may provide a highly streamlined method of DNA extraction for fresh, ethanol-preserved, and young, dried specimens, especially when adapted for high-throughput, robotic systems.     

A pressure cooking-based DNA extraction from archival formalin-fixed, paraffin-embedded tissue

As emerging novel DNA-based methodologies are adopted, nucleic acid-based assays depend critically on the quality and quantity of extracted DNA. Formalin-fixed, paraffin embedded (FFPE) tissue samples provide an invaluable resource for subsequent molecular studies of clinical phenotypes, but high-quality DNA extraction from archival FFPE tissue specimens remains complex and time-consuming. To address this challenge, we have developed a reliable rapid DNA extraction method for FFPE tissue specimens. It is based on deparaffinization at high temperature coupled with relieving crosslink in a pressure cooker. The DNA yield by this rapid method resulted in an average 1.8-fold increase in comparison with the commercial kit and OD 260/280 ratios between 1.87 and 1.95. The DNA obtained by the rapid method was suitable for methylation analyses in colon cancer patients. These data suggest that this new DNA extraction method coupled with methylation-specific polymerase chain reaction can be used for epigenetic studies with the advantages of rapidity and high quality and may contribute to the development of biomarkers in clinical studies.