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1.
对野生动物进行分子水平研究时,在取材上经常存在野外找不到动物或得到的材料提取不到足够的DNA等困难。但随着PCR技术的产生和DNA提取技术的进步,研究材料也不再局限于新鲜的组织样本,从而取样的范围扩大到微量的血液、单根的毛发、羽毛、指甲、唾液、粪便、骨骼,甚至古代化石等样品,研究范围随之也不断扩大。 相似文献
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太白红杉3种不同材料总DNA的提取 总被引:6,自引:0,他引:6
高质量DNA的获得是进行各项遗传操作的基础,提取DNA的方法、材料因不同的植物而异。我们用改良的CTAB法,对太白红杉的3种不同材料的基因组总DNA进行了提取,均成功获得了适于RAPD分析的总DNA。结果表明,提取太白红杉DNA并对其进行研究,幼苗是较佳材料,愈伤组织也是较好的可试材料,而针叶则相对较差。 相似文献
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野老鹳草DNA的提取方法及RAPD分析 总被引:1,自引:0,他引:1
目的:以野老鹳草(Geranium carolinianum L.)的茎、叶为材料,研究野老鹳草DNA的提取方法及RAPD的分析。方法:采用CATB法、高盐低pH法和SDS法分别提取野老鹳草的基因纽DNA,并对3种方法进行了一些改进。通过RAPD分析所提取的DNA,比较所用的提取方法。结果:比较DNA产量、质量等,确定了高盐低pH法较佳。结论:干燥的材料和新鲜的材料均可提取得到DNA,高盐低pH法提取的效果优于CTAB和SDS法。 相似文献
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对影响哺乳动物粪便DNA提取相关因素的探讨 总被引:4,自引:0,他引:4
从动物粪便中提取DNA是一种优秀的非损伤性取样方法,然而在实际操作中成功提取高质量粪便DNA却是一件不太容易的事情.粪便DNA的获取不仅与提取方法有关,还受到样品采集、保存、二次取样、预处理等相关环节的影响,对其中任一环节的忽视都会导致试验达不到理想效果.综合国内外具有代表性的哺乳动物粪便DNA提取技术,对有关环节进行了详细的评述,并对PCR扩增中的常见问题进行了分析和讨论. 相似文献
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海洋沉积物柱状样有孔虫古DNA是近年来国际新兴的研究技术,对于解释海洋全球变化可以获取到传统形态学检获不到的遗传信息,并对其进行比较和补充。目前国际上有孔虫古DNA的研究主要开展于深海和极地等有利于DNA保存的环境,但对于近海陆架海域尚无相关有效的技术研究。为了探索适合提取和扩增陆架浅海环境沉积物柱状样中的有孔虫古DNA的方法,本实验以采自山东半岛附近的黄海沉积物柱状样为研究对象,通过改进DNA提取过程的涡旋震荡时间和洗脱液体积、比较不同的PCR扩增条件和引物对,对陆架浅海地区沉积物柱状样中有孔虫古DNA提取和PCR扩增的方法进行了探索。借助ImageJ软件对PCR产物的凝胶电泳图像进行了定量分析与比较。研究结果显示,延长涡旋震荡时间和减少洗脱液体积可以提高对海洋沉积物环境总古DNA的提取效能,使用引物对s14F0和s15以及优化后的PCR条件能成功扩增陆架浅海环境沉积物中的有孔虫古DNA。本文探索了适用于陆架浅海环境沉积物柱状样的有孔虫古DNA的研究技术,可为古海洋和古环境研究提供新的研究思路。 相似文献
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《微体古生物学报》2022,(1)
海洋沉积物柱状样有孔虫古DNA是近年来国际新兴的研究技术,对于解释海洋全球变化可以获取到传统形态学检获不到的遗传信息,并对其进行比较和补充。目前国际上有孔虫古DNA的研究主要开展于深海和极地等有利于DNA保存的环境,但对于近海陆架海域尚无相关有效的技术研究。为了探索适合提取和扩增陆架浅海环境沉积物柱状样中的有孔虫古DNA的方法,本实验以采自山东半岛附近的黄海沉积物柱状样为研究对象,通过改进DNA提取过程的涡旋震荡时间和洗脱液体积、比较不同的PCR扩增条件和引物对,对陆架浅海地区沉积物柱状样中有孔虫古DNA提取和PCR扩增的方法进行了探索。借助ImageJ软件对PCR产物的凝胶电泳图像进行了定量分析与比较。研究结果显示,延长涡旋震荡时间和减少洗脱液体积可以提高对海洋沉积物环境总古DNA的提取效能,使用引物对s14F0和s15以及优化后的PCR条件能成功扩增陆架浅海环境沉积物中的有孔虫古DNA。本文探索了适用于陆架浅海环境沉积物柱状样的有孔虫古DNA的研究技术,可为古海洋和古环境研究提供新的研究思路。 相似文献
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苯酚是进行高质量植物DNA提取常用的试剂,但苯酚对患病材料中病原检测及后续分子生物学试验的影响尚未见报道。本试验利用苯酚对健康和患枣疯病的枣树叶片中DNA进行提取,并进行了后续的病原检测和AFLP分析,结果表明(1)苯酚抽提获得的DNA质量及产率明显高于对照;(2)苯酚抽提处理对枣疯病病原的检测有直接影响,当病原含量较低时,加苯酚抽提后的DNA检测不到病原;(3)苯酚抽提处理与否获得的DNA对后续AFLP分析的扩增效果没有明显影响;但两者出现了有规律的差异条带,经分析此差异应该是由材料中有无病原引起的。本试验结果为寄主材料中病原检测及寄主与病原之间的互作等研究提供了有益参考。 相似文献
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一种高效的哺乳动物粪便DNA提取通用方法 总被引:2,自引:0,他引:2
以粪便为材料提取动物DNA进行动物保护遗传学和分子生态学研究的关键是能否提取到高质量的粪便DNA.然而提取方法通用性不好和产物质量不高等问题阻碍了粪便DNA分析技术的推广.本文介绍的改进型十六烷基三甲基溴化铵提取法可广泛适用于各食性哺乳动物粪便DNA提取,在11种不同食性动物的粪便DNA提取实验中验证了它的可靠性和通用性.本方法成本低廉(3元/样),用实验室常规试剂即可完成粪便DNA提取,其产物纯度高于专用试剂盒QIAamp DNA Stool Kit,在拥有超过专业试剂盒提取效果的同时尽可能的降低了实验成本,有利于粪便DNA技术的推广. 相似文献
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A water soluble extract was prepared from the lymph nodes of rats of different age groups. This extract, when added to human peripheral blood leucocyte cultures, inhibits DNA synthesis and mitosis when added in different concentrations for varying lengths of time in culture. Also, it was observed that the extract from old rats was not as effective as that from younger animals. 相似文献
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Rodrigo R. Domingues Domingos Garrone-Neto Alexandre W. S. Hilsdorf Otto B. F. Gadig 《Journal of fish biology》2019,94(3):512-516
In this study we tested the use of mucus from five species of Neotropical marine batoid elasmobranchs to extract genomic DNA for barcoding and phylogenetic analysis. The DNA from all individuals sampled was successfully amplified and sequenced for molecular barcode, allowing 99–100% accuracy to the species level. This method proved to provide reliable and good-quality DNA for barcoding and phylogenetic analysis of Neotropical elasmobranchs, through rapid handling and with low disturbance to animals. 相似文献
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Collecting and storing biological material from wild animals in a way that does not deteriorate DNA quality for subsequent analyses is instrumental for research in ecology and evolution. Our aims were to gather reports on the effectiveness of methods commonly used by researchers for the field collection and long‐term storage of blood samples and DNA extracts from wild birds. Personal experiences were collected with an online survey targeted specifically at researchers sampling wild birds. Many researchers experienced problems with blood sample storage but not with DNA extract storage. Storage issues generated problems with obtaining adequate DNA quality and sufficient DNA quantity for the targeted molecular analyses but were not related to season of blood sampling, access to equipment, transporting samples, temperature, and method of blood storage. Final DNA quality and quantity were also not affected by storage time before DNA extraction or the methods used to extract DNA. We discuss practical aspects of field collection and storage and provide some general recommendations, with a list of pros and cons of different preservation methods of avian blood samples and DNA extracts. 相似文献
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对于自然环境中的或长期保存的动物标本,由于保存环境不良或保存时间过长,DNA提取的难度较大。受标本保存时间和损害程度等因素影响,导致实验结果的不稳定性加强,对于同一标本需要反复实验。为了提高DNA提取效率,节省实验成本,现对陈旧损坏标本的DNA提取方法进行综述。 相似文献
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Campos-da-Paz M Pereira LO Bicalho LS Dórea JG Poças-Fonseca MJ Santos Mde F 《Mutation research》2008,652(2):158-163
The consumption of bracken-fern (Pteridium aquilinum) as food is associated with a high incidence of cancer in humans and animals. Thus far, the carcinogenic effects of bracken-fern consumption could be related to chromosome aberrations verified in animal and in human peripheral lymphocytes. We tested the in vitro effects of vitamin C (10 and 100 microg/ml) on the reversibility of DNA damage caused by bracken-fern on human submandibular gland (HSG) cells and on oral epithelium cells (OSCC-3) previously exposed to bracken-fern extract. DNA damage (i.e. nuclei with increased levels of DNA migration) was determined by comet assay, cell morphology was evaluated by light microscopy and cellular degeneration was assessed by the acridine orange/ethidium bromide fluorescent-dyeing test. Results showed that vitamin C alone did not reduce DNA damage caused by bracken-fern in HSG and OSSC-3 cells. However, at a higher concentration (100 microg/ml), vitamin C induced DNA damage in both cell lines. Moreover, vitamin C (10 and 100 microg/ml) together with bracken-fern extract showed synergistic effects on the frequency of DNA damage in HSG cells. In addition, cells treated with bracken-fern extract or vitamin C alone, or with their association, showed apoptosis morphological features, such as chromatin condensation, cytoplasmic volume loss, changes in membrane symmetry and the appearance of vacuoles; these alterations were observed in both cell lines. These results demonstrate that bracken-fern extract was cytotoxic to HSG and OSCC-3 cells, causing cell death by apoptosis, and that vitamin was not able to revert these effects. 相似文献
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Unilateral nephrectomy (uNX) in mice is followed by a transitory increase in cell proliferation in the remaining kidney. To examine whether this response could be related to a negative feedback control of kidney epithelial cell renewal, water extracts were made of kidney homogenate. Five mg freeze-dried extract was injected 18 h post-operatively, and the animals were sacrificed at intervals during the following 54 h. The mitotic rate and the incorporation of tritiated thymidine (3H-TdR) into DNA were measured in the remaining kidney. The results show that the kidney extract reduces both the mitotic rate and the incorporation of 3H-TdR into DNA. In the tubular epithelium in the kidney, the strongest inhibitory effect was found by injecting the extract at 18 or 39 h postoperatively. 相似文献
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Adult rats with normal or X-irradiated testes were used in an experiment to test the possible existence of a chalone in the testis. On the 11th day following irradiation, i.e. as the type A spermatogonia proliferated actively to restore the partially destroyed spermatogonial population, the animals with irradiated testes were subdivided into three groups. Rats of the first group were injected intraperitoneally with a saline extract of normal adult rat testes. Animals of the second group were injected with an equal amount of physiological saline while the rats of the third group received equivalent injections of a saline liver extract. Two additional groups of rats with non-irradiated testes, injected with the testicular extract or saline solution, served as controls. Following the last injection all animals were injected with 3H-thymidine and sacrificed. From each animal one testis was used to determine the specific radioactivity of its DNA, the other testis was processed for radioautography. The testicular extract produced a significant decrease in uptake of radioactivity by the irradiated testes. There was no difference in the radioactivity uptake by the testes of non-irradiated rats. Correspondingly the labeling index of type A spermatogonia was significantly lower in animals of the first group than in the other two groups of animals with irradiated testes. However, there was no difference in the labeling indices of Intermediate and type B spermatogonia or of preleptotene spermatocytes in the animals receiving the extracts or the saline solution. In animals with non-irradiated testes there was no difference in the labeling indices of type A or other types of spermatogonia or of spermatocytes. These data were taken to indicate that a saline extract of normal adult testes contains a substance that can inhibit specifically the proliferation of type A spermatogonia during the repair phase of the spermatogonial population following irradiation. This substance was tentatively considered as a spermatogonial chalone. 相似文献
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OA Lemos JC Sanches IE Silva ML Silva AH Vinhólis MA Felix RA Santos AO Cecchi 《Genetics and molecular biology》2012,35(2):498-502
Tabebuia sp. is native to tropical rain forests throughout Central and South America. Although the biological and pharmacological effects of bark extracts have been intensely studied, little is known on the extract obtained from the flower. Herein, the genotoxic potential of a flower extract from T. impetiginosa ("ipê roxo") on the blood and liver cells of Wistar rats was evaluated. Experimental procedures involved only male animals. Graduated concentrations of the extract, viz., 100, 300 and 500 mg kg(-1) of body weight, were gavage-administered and 24 h latter cells were collected and processed for analysis. With the exception of the 100 mg kg(-1) dose, a significant increase in DNA damage was noted, when compared with a negative control group. Although the genotoxic potential of this extract was higher in liver cells, the response in both tissues was related to dose-dependency. Even though DNA damage can be corrected before conversion into mutations, further study is recommended to arrive at a better understanding of incurred biological effects. 相似文献
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Mariana Campos-da-Paz Luciana O. Pereira Leandro Santos Bicalho Jos G. Drea Marcio J. Poas-Fonseca Maria de Ftima M. Almeida Santos 《Mutation Research - Genetic Toxicology and Environmental Mutagenesis》2008,652(2):158-163
he consumption of bracken-fern (Pteridium aquilinum) as food is associated with a high incidence of cancer in humans and animals. Thus far, the carcinogenic effects of bracken-fern consumption could be related to chromosome aberrations verified in animal and in human peripheral lymphocytes. We tested the in vitro effects of vitamin C (10 and 100 μg/ml) on the reversibility of DNA damage caused by bracken-fern on human submandibular gland (HSG) cells and on oral epithelium cells (OSCC-3) previously exposed to bracken-fern extract. DNA damage (i.e. nuclei with increased levels of DNA migration) was determined by comet assay, cell morphology was evaluated by light microscopy and cellular degeneration was assessed by the acridine orange/ethidium bromide fluorescent-dyeing test. Results showed that vitamin C alone did not reduce DNA damage caused by bracken-fern in HSG and OSSC-3 cells. However, at a higher concentration (100 μg/ml), vitamin C induced DNA damage in both cell lines. Moreover, vitamin C (10 and 100 μg/ml) together with bracken-fern extract showed synergistic effects on the frequency of DNA damage in HSG cells. In addition, cells treated with bracken-fern extract or vitamin C alone, or with their association, showed apoptosis morphological features, such as chromatin condensation, cytoplasmic volume loss, changes in membrane symmetry and the appearance of vacuoles; these alterations were observed in both cell lines. These results demonstrate that bracken-fern extract was cytotoxic to HSG and OSCC-3 cells, causing cell death by apoptosis, and that vitamin was not able to revert these effects. 相似文献
20.
Hwang IK Yoo KY Kim DS Jeong YK Kim JD Shin HK Lim SS Yoo ID Kang TC Kim DW Moon WK Won MH 《Life sciences》2004,75(16):1989-2001
Grape seed extract (GSE) possess cardioprotective abilities by functioning as in vivo antioxidants and by virtue of their ability to directly scavenge ROS including hydroxyl and peroxyl radicals. In the present study, we investigated the neuroprotective effects of grape seed extract (GSE) in the gerbil hippocampus after 5 min transient forebrain ischemia. Neuronal cell density in GSE-treated ischemic animals was significantly increased as compared with vehicle-treated ischemic animals 4 days after ischemic insult. In the GSE-treated groups, about 60% of pyramidal cells of the sham-operated group were stained with cresyl violet 4 days after ischemic insult. In this study, we found that GSE had neuroprotective effects on neuronal injury by inhibiting DNA damage in the CA1 region after ischemia. In vehicle-treated groups, 8-hydroxy-2'-deoxyguanosine (8-OHdG) immunoreactivity was significantly changed time-dependently, whereas the immunoreactivity in the GSE-treated group was similar to the sham-operated group. In addition, we confirmed that astrocytes and microglia did not show significant activation in the CA1 region 4 days after ischemia-reperfusion, because many CA1 pyramidal cells were not damaged. Therefore, these results suggest that GSE can protect ischemic neuronal damage by inhibiting DNA damage after transient forebrain ischemia. 相似文献