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Evaluation of six methods for total dna extraction from gut earthworms for microbial community diversity and metagenomics. Repositório Alice
ESTEVES, E. D.; BROWN, G. G.; PEDROSA, F. de O.; SOUZA, E. M. de; CHUBATSU, L. S..
2015
Tipo: Resumo em anais de congresso (ALICE) Palavras-chave: Minhoca; Extração de DNA; Método; Earthworm; DNA extraction.
Ano: 2015 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1029457
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Extração de DNA a partir de coágulos sanguíneos bovinos. Infoteca-e
BRITO, L. G.; OLIVEIRA, M. C. de S.; MOURA, M. M. da F.; SILVA NETTO, F. G. da S.; CAVALCANTE, F. A.; MARIM, A. D.; SOUZA, G. C. R. de; SILVA, J. L. da.
Foi otimizada uma metodologia de extração de DNA genômico a partir de coágulo sanguíneo. Amostras de sangue com e sem anticoagulante EDTA para a obtenção dos coágulos sanguíneos foram coletadas em bovinos nos estados de Rondônia e Acre para pesquisa de Anaplasma marginale. Para a extração de DNA a partir do coágulo sanguíneo testaram-se dois tipos de tecido de nylon: no tratamento 1, o tecido utilizado apresentava diâmetro dos poros de 200 µm e no tratamento 2, tecido com poros de diâmetro de 1.200 µm. As amostras de sangue com anticoagulante representaram o tratamento 3, sendo este considerado como tratamento controle. Após a quebra mecânica, os coágulos sofreram centrifugação a 7.000 RPM por 15 minutos passando através das malhas dos tecidos utilizados...
Tipo: Boletim de Pesquisa e Desenvolvimento (INFOTECA-E) Palavras-chave: Extração de DNA; Coágulo sanguíneo; Epidemiologia molecular; Doenças de bovinos; DNA extraction; Blood clot; Molecular epidemiology; Bovine diseases.
Ano: 2006 URL: http://www.infoteca.cnptia.embrapa.br/handle/doc/710703
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Extraction of high-quality host DNA from feces and regurgitated seeds: a useful tool for vertebrate ecological studies Biol. Res.
MARRERO,PATRICIA; FREGEL,ROSA; CABRERA,VICENTE M; NOGALES,MANUEL.
DNA extraction methods for genotyping non-invasive samples have led to great advances in molecular research for ecological studies, and have been particularly useful for analyzing threatened species. However, scarce amounts of fragmented DNA and the presence of Taq polymerase inhibitors in non-invasive samples are potential problems for subsequent PCR amplifications. In this study we describe a novel technique for extracting DNA from alimentary tract cells found on external surfaces of feces and regurgitated seeds. The presence of contaminants and inhibitors is minimized and samples are preserved intact for use in other ecological research (e.g. trophic studies). The amplification efficiency and purity of the extracted DNA from feces were significantly...
Tipo: Journal article Palavras-chave: DNA extraction; Ecological studies; Guanidine thiocyanate; Non-invasive method; Pigeons; Seed dispersal.
Ano: 2009 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602009000200002
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Individual genomic DNA isolation of Entomogenous nematodes: the Deladenus study case. Repositório Alice
SCHUHLI, G. S. e; PENTEADO, S. do R. C.; KESTRING, D. R.; STEILL, G. J.; THOMAS, M. C..
Estudos populacionais envolvem a extração de DNA de um único indivíduo para permitir a genotipagem. No caso de alguns nematoides parasíticos, suas dimensões reduzidas e o grande número de indivíduos por infestação tornam a genotipagem difícil. Com o objetivo de desenvolver um protocolo, foram feitos ajustes em métodos já disponíveis, visando melhor ganho em pureza e concentração de DNA genômico. Espécimes foram digeridos em tampão de digestão de verme e submetidos à amplificação por PCR, como prova de conceito. Foi possível a obtenção de boa quantidade e concentração de DNA de indivíduos. A qualidade foi suficiente para garantir o sequenciamento da região ITS1.
Tipo: Nota Técnica/Nota Científica (ALICE) Palavras-chave: Sirex noctilio; Vespa da madeira; Nematoide; Extração de DNA; Protocolo; Woodwasp; DNA extraction.
Ano: 2012 URL: http://www.alice.cnptia.embrapa.br/handle/doc/945902
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Isolation of high quality DNA: a protocol combining “rennet” and glass milk Electron. J. Biotechnol.
Valter de Oliveira,Luiz Felipe; Wallau,Gabriel da Luz; Silva Loreto,Elgion Lucio.
High quality DNA is essential for many molecular biology techniques. However, the reagents used for that purpose usually are expensive and/or cause a high environmental impact. Here, we describe two alternative protocols that use inexpensive reagents and are not hazardous to the environment. The first protocol utilizes the enzyme chymosin, normally used as “rennet” in cheese production and which is easily obtained on the commercial market. The second protocol uses “rennet DNA extraction protocol” combined with the DNA binding capacity of glass powder (glass milk), which can easily be “home made”. The first protocol is used when a high yield of DNA is needed, whereas the second protocol is used for...
Tipo: Journal article Palavras-chave: Chymosin; DNA extraction; Glass milk; Rennet; Rennin; Silica.
Ano: 2009 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582009000200011
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Método não-invasivo na obtenção de DNA de búfalos - DOI: 10.4025/actascianimsci.v30i4.4839 Animal Sciences
Ribeiro, Juliana Maria; UEM; Gasparino, Eliane; UEM; Marques, Débora Sommer; UEM; Luizetti, Fabiane; UEM; Soares, Maria Amélia Menck; UEM; Zeoula, Lúcia Maria; UEM.
O objetivo do trabalho foi comparar dois diferentes protocolos de extração de DNA de pelos de búfalos (Bubalus bubalis) e comparar três regiões de coleta de material (nuca, paleta direita e testa). Foram utilizados quatro búfalos com três repetições por animal e por região. No protocolo 1, foi utilizada a técnica do fenol-clorofórmio e no protocolo 2, a técnica de extração com CTAB. O protocolo 2 apresentou maior média de concentração de DNA para as amostras de pelos. Em relação ao local de retirada dos pelos, não foram encontradas diferenças significativas, porém nota-se que a região da testa dos animais apresentou maior concentração de DNA quando extraído com CTAB. Com relação à praticidade de utilização dos dois métodos avaliados, o protocolo 2, além de...
Tipo: Análise laboratorial Palavras-chave: 5.05.04.00-2 Reprodução Animal CTAB; Extração de DNA; Fenol; Pelos. Genética e Melhoramento dos Animais Domésticos CTAB; DNA extraction; Phenol; Fur..
Ano: 2009 URL: http://periodicos.uem.br/ojs/index.php/ActaSciAnimSci/article/view/4839
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Microrganismos associados às cigarrinhas vetoras de Xylella fastidiosa. Repositório Alice
SOUZA, A. C. de; FONSECA, P. da; DORNELES JUNIOR, J.; SANTOS, E. R. dos; PRADO, S. de S..
As cigarrinhas são insetos sugadores da família Cicadellidae que se alimentam principalmente nos vasos do xilema de plantas de citros, café e ameixa e transmitem a bactéria Xylella fastidiosa para estas plantas. Os principais sintomas na planta incluem amarelecimento e redução do tamanho de folhas, encurtamento de internódios, abscisão foliar e seca de ramos. Além de patógenos de plantas, os insetos se encontram associados a simbiontes, os quais podem ou não ser encontrados em especificas células nos insetos, proporcionando um benefício mutuo na maioria dos casos. O presente estudo teve como objetivo detectar a presença do fitopatógeno X. fastidiosa e dos simbiontes Candidatus Sulcia muelleri e Betaproteobacteria em três espécies de cigarrinhas, coletadas...
Tipo: Artigo em anais de congresso (ALICE) Palavras-chave: Cicadellidae; Simbionte; Bactéria fitopatógenica; Extração de DNA; DNA extraction; Cigarrinha; Bactéria; Cicadellidae; Symbionts; Plant pathogenic bacteria.
Ano: 2017 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1084323
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Protocol for extraction of genomic DNA from swine solid tissues Genet. Mol. Biol.
Biase,Fernando Henrique; Franco,Maurício Machaim; Goulart,Luiz Ricardo; Antunes,Robson Carlos.
Molecular diagnostics are performed by using DNA from different body tissues. However, it is necessary to obtain genomic DNA of good quality. Due to the impossibility of collecting blood from slaughtered animals, DNA extraction from solid tissues is necessary. The objective of this study was to describe a protocol of DNA extraction from swine skin, adipose, brain, liver, kidney and muscle tissues. We obtained high molecular weight DNA of good quality, shown by agarose gel and amplification of two DNA fragments, 605bp and 891pb, by PCR. Spectrophotometric analysis of DNA concentration showed variation among the DNA from different tissues, with the liver and adipose tissues presenting the greatest and the smallest concentration, respectively. The described...
Tipo: Info:eu-repo/semantics/other Palavras-chave: DNA extraction; Swine tissues.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572002000300011
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Rapid yeast DNA extraction by boiling and freeze-thawing without using chemical reagents and DNA purification BABT
Silva,Gildo Almeida da; Bernardi,Taís Letícia; Schaker,Patrícia Dayane Carvalho; Menegotto,Morgana; Valente,Patricia.
The purpose of this work was to study a rapid yeast DNA extraction by boiling and freeze-thawing processes without using chemical reagents or any purification procedures, to obtain a high grade PCR-product. A specific DNA fragment of the 18S region of Dekkera bruxellensis and Saccharomyces cerevisiae was chosen. The described boiling and freeze-thawing protocols generated the PCR-grade product preparations and could be used to process many samples. The amplification of the fragments could be observed after 30 and 35 cycles. These processes of extraction without using any kind of chemical reagents, especial water, and purification procedures proved to be efficient, reproducible, simple, fast, and inexpensive.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Boiling; Freeze-thawing; DNA extraction; Dekkera bruxellensis; Saccharomyces cerevisiae.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000200020
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Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea mays Ciencia e Investigación Agraria
Barra,Macarena; Salazar,Erika; Beltrán,María; Sagredo,Boris.
The extraction of high-quality DNA is essential for studies conducted at the molecular level in species with an abundance of contaminants in their tissues, such as some landraces of potato and maize, in which it is difficult to extract good-quality genomic DNA. Compounds such as polyphenols interfere with the amplification of DNA during polymerase chain reaction (PCR), which makes it difficult to conduct PCR-based studies ofmolecular markers. This article describes a simple and robust protocol for DNA isolation that was applied to potato and maize landraces and resulted in a high DNA concentration with excellent purity and quality. This method is based on the method of Krizman et al. (2006) with some modifications and was tested on potato leaves and young...
Tipo: Journal article Palavras-chave: Activated charcoal; DNA extraction; Genomic DNA; Polyphenols.
Ano: 2012 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0718-16202012000300019
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Simple method to prepare DNA templates from a slice of peanut cotyledonary tissue for Polymerase Chain Reaction Electron. J. Biotechnol.
Yu,Shu Tao; Wang,Chuan Tang; Yu,Shan Lin; Wang,Xiu Zhen; Tang,Yue Yi; Chen,Dian Xu; Zhang,Jian Cheng.
An efficient DNA extraction method was developed for peanut seed, where only 3-5 mg cotyledonary tissue was enough for more than 50 PCR reactions with a reaction volume of 15 μl. Both low copy number and high copy number DNA sequences were successfully amplified. Processing one seed sample only took about half an hour. Sampling had no significant effects on germination and development. The DNA extraction method makes it possible to identify transformants and conduct molecular marker studies prior to sowing, and thus may greatly hasten research progress.
Tipo: Journal article Palavras-chave: Cotyledonary tissue; DNA extraction; Groundnut; PCR; Peanut.
Ano: 2010 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000400012
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Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis Genet. Mol. Biol.
Scatena,Mário Pinzan; Morielle-Versute,Eliana.
To establish a technique which minimized the effects of fixation on the extraction of DNA from formalin-fixed tissues preserved in scientific collections we extracted DNA samples from fixed tissues using different methods and evaluated the effect of the different procedures on PCR and sequencing analysis. We investigated muscle and liver tissues from museum specimens of five species of fruit-eating (frugivorous) bats of the Neotropical genus Artibeus (Chiroptera, Phyllostomidae): A. fimbriatus, A. lituratus, A. jamaicensis, A. obscurus, and A. planirostris. The results indicated that treatment of tissues in buffered solutions at neutral pH and about 37 °C for at least four days improves the quality and quantity of extracted DNA and the quality of the...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Archival specimens; Chiroptera; DNA extraction; Formalin-fixed.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572008000100027
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The use of museum specimens as source of DNA Naturalis
Cibois, A..
Tipo: Article / Letter to the editor Palavras-chave: Aves; Museum specimens; DNA extraction; 42.83.
Ano: 2005 URL: http://www.repository.naturalis.nl/record/210819
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Urine as a promising sample for Leishmania DNA extraction in the diagnosis of visceral leishmaniasis - a review BJID
Bezerra,Gilberto Silva Nunes; Barbosa Júnior,Walter Lins; Silva,Elis Dionísio da; Leal,Nilma Cintra; Medeiros,Zulma Maria de.
ABSTRACT Visceral leishmaniasis is a serious and debilitating infection with high fatality rate in tropical and subtropical countries. As clinical symptoms of visceral leishmaniasis are not so specific, confirmatory diagnostic methods with high sensitivity and specificity are needed. Noninvasive methods have been developed using urine as a clinical sample for visceral leishmaniasis diagnosis. In fact, there is a clear correlation between kidney impairment and Leishmania DNA in urine. However, it has been proved that Leishmania nucleic acid may also be isolated from patients without any sign of renal involvement. Even though urine has become a promissing biological sample, it is still not widely used due to several issues, such as (i) incomprehension of the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Visceral leishmaniasis; Renal involvement; Diagnosis; Urine sample; DNA extraction.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702019000200111
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Use of a cysteine proteinase from Carica candamarcensis as a protective agent during DNA extraction BJMBR
Genelhu,M.S.; Zanini,M.S.; Veloso,I.F.; Carneiro,A.M.D.; Lopes,M.T.P.; Salas,C.E..
We describe the use of a plant cysteine proteinase isolated from latex of Carica candamarcensis as a protective agent during isolation of bacterial DNA following growth in culture of these cells. Between 100 to 720 units of proteinase (1 µg = 6 units) afforded good DNA protection when incubated with various kinds of microorganisms. Agarose gel electrophoresis showed that the resulting DNA was similar in size to DNA preparations obtained by treatment with proteinase K. The viability of the resulting material was checked by PCR amplification using species-specific primers. After standing at room temperature (25oC) for 35 days, the enzyme lost 10% of its initial activity. The enzyme stability and good yield of DNA suggest the use of this proteinase as an...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Cysteine proteinase; Proteinase K; C. candamarcensis; DNA extraction.
Ano: 1998 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1998000900005
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Use of real-time PCR to evaluate two DNA extraction methods from food Ciênc. Tecnol. Aliment.
Branquinho,Maria Regina; Ferreira,Renata Trotta Barroso; Cardarelli-Leite,Paola.
The DNA extraction is a critical step in Genetically Modified Organisms analysis based on real-time PCR. In this study, the CTAB and DNeasy methods provided good quality and quantity of DNA from the texturized soy protein, infant formula, and soy milk samples. Concerning the Certified Reference Material consisting of 5% Roundup Ready® soybean, neither method yielded DNA of good quality. However, the dilution test applied in the CTAB extracts showed no interference of inhibitory substances. The PCR efficiencies of lectin target amplification were not statistically different, and the coefficients of correlation (R²) demonstrated high degree of correlation between the copy numbers and the threshold cycle (Ct) values. ANOVA showed suitable adjustment of the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: DNA extraction; Genetically modified organisms; GMO; Food analysis; Real-time PCR; Linearity.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612012000100017
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การสกัดดีเอ็นเอจากข้าวโพดในระดับเล็กสำหรับทำพีซีอาร์ Thai Agricultural
Wanchai Yenpetch; Chaba Jampatong; Sanon Suksatan.
This method for instant DNA extraction from fresh corn leaf based on a simple DNA extraction with CTAB modified from Doyle and Doyle (1987) in small scale. After extraction steps, the extracted DNA was checked the quality and quantity by using a spectrophotometer to measure the absorbance at A260/A280. The result showed the feasibility of this method which was confirmed by using PCR method. The PCR product showed an expected band. Without the addition of the phenol or 2-beta-mercaptoethanol, this method could properly use in routine laboratory where DNA is used for PCR detection. In addition, this method requires less plant tissue to extract that is useful for other plant experiments.
Palavras-chave: Corn; Leaf; DNA extraction; PCR; ข้าวโพด; ใบสด; ดีเอ็นเอ; การสกัด; พีซีอาร์; คุณภาพ; สเปคโตรโฟโตมิเตอร์; อัตราการดูดกลืนแสง.
Ano: 2010 URL: http://anchan.lib.ku.ac.th/agnet/handle/001/4690
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木瓜基因組 DNA 快速萃取技術之開發與轉基因木瓜檢測技術之建立 Taiwan Agricultural Research Institute
李文立; 王德男; Wen-Li Lee; Der-Nan Wang.
[[abstract]]自1975年起,台灣木瓜因遭受病毒病的危害而使木瓜產業衰退,雖然採用網室栽培可以防杜病毒病危害,但也會使栽培成本增加,成為栽培上的一項限制。近年來,生物技術的快速發展及轉基因作物的商業化,利用基因工程育成之抗病毒病轉基因木瓜分別在夏威夷、台灣及泰國等地被開發。然而生物安全性議題備受重視,世界先進國家均訂定轉基因作物之生物安全評估與管理制度。因此檢測技術的開發在世界各國均受到重視,又因為國際貿易上,農產品的輸出入為重要一環。快速而正確的檢測方法將會是一項重要關鍵。目前在轉基因作物檢測上仍以PCR法為最常用之方式,然而檢測前的DNA萃取工作往往耗時甚久(一般約2-8小時),成為檢測上主要的成本支出與限制,本研究利用不同比例之界面活性劑及緩衝液作為萃取液,可以大幅減少基因組DNA萃取時間,增進工作效率,此一快速萃取方式也同時可以用在不同種類的果樹及蔬菜作物上,是一種方便且廣效的創新 方法。另外,依照目前的檢測流程在檢測工作上,必須同時分別進行外源基因(轉 殖入的基因)檢測以判定是否為轉基因木瓜及papain基因之擴增,以確認實驗是 否有誤差。分別檢測的方式不僅,在人力、物力上需耗費許多的檢測成本,本研 究中也發展多重PCR檢測方式,在同一試管中進行多引子對同時偵測外源基因及 papain基因,不僅可以去除偽陰性也同時可節省檢測時間與經費。 Since 1975, the industry of papaya was series damage by papaya ring sport virus in Taiwan. Although the net house culture techniques are a good practice to protect papaya avoid...
Palavras-chave: 木瓜; DNA萃取; 快速; 轉基因; 檢測; 聚合酵素鍊鎖反應 台灣木瓜產業發展研討會專刊 papaya; DNA extraction; Rapid; Transgenic; Detection; PCR [[classification]]15.
Ano: 2006
Registros recuperados: 38
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