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Registros recuperados: 38
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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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Comparison of preservation methods of Atta spp. (Hymenoptera: Formicidae) for RAPD analysis Anais da SEB
Carvalho,Alfredo O. R.; Vieira,Luiz G. E..
High quality DNA for molecular studies can be easily extracted from fresh specimens. However, live samples are difficult to keep for long periods thus making their preservation a serious problem, specially when they are collected and transported from remote locations. In order to establish an efficient method to preserve Atta spp. (leaf-cutting ants) for RAPD analysis, six different storage methods were examined: 1) -70°C; 2) 95% ethanol at -20°C; 3) 95% ethanol at 4°C; 4) 95% ethanol at room temperature; 5) silica gel at room temperature; and 6) buffer (0.25 M EDTA, 2.5% SDS, 0.5 M Tris-HCl, pH 9.2) at room temperature. DNA was extracted (Cheung et al., 1993 - modified) and examined after 90, 210 and 360 days of storage. Freshly killed specimens were used...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Insecta; Leaf-cutting ants; DNA extraction; Storage conditions.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0301-80592000000300011
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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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Enhancement of soil DNA extraction by the use of a hand held mixer BJM
Costa,Jefferson Luis da Silva; Oliveira,Virgínia Carla de.
The efficiency of soil DNA extraction using a bead beater homogenizer (Biospec Products - Germany) was compared to that obtained with a hand held mixer (Moulinex - Brazil). The hand held mixer costs 100 times less and extracted seven times more crude DNA than the bead beater.
Tipo: Info:eu-repo/semantics/other Palavras-chave: Molecular tools; DNA extraction.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000400004
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Direct RAPD evaluation of bacteria without conventional DNA extraction BABT
Araújo,Welington Luiz; Angellis,Derlene Attili de; Azevedo,João Lúcio.
The present work reports successful DNA amplification of Pantoea agglomerans and Bacillus pumilus through Random Amplified Polymorphic DNA (RAPD). For this, template DNA was obtained without conventional DNA extraction. The procedure was as follows: cultures grown for 20 hours in 5 mL LB medium were centrifuged and the resulting preparation was suspended in TE buffer. After boiling, the cell suspension was diluted and 2.0 µl were used in reactions of 15 µl. The results showed no significant differences among the RAPD profile of the PCR reactions derived from the boiling and phenol extraction methods, suggesting the utilization of this method for genetic population analysis.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bacteria; Boiling method; DNA extraction; RAPD.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132004000300006
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Comparison of three methods of DNA extraction from cold-smoked salmon and impact of physical treatments ArchiMer
Giacomazzi, Sophie; Leroi, Francoise; Joffraud, Jean-jacques.
Aims: To compare three bacterial DNA extraction procedures on cold-smoked salmon (CSS) and assess the impact on their efficiency of two physical treatments of the food matrix, ionizing irradiation and freezing. Methods and Results: As molecular methods for bacterial detection have become an important analytical tool, we compared bacterial DNA extraction procedures on CSS. Working with frozen and irradiated CSS, we obtained negative responses from samples known to be highly contaminated. Thus, we decided to study the impact of these two physical treatments on bacterial DNA extraction procedures. The efficiency of bacterial DNA extraction directly from the fish matrix suspension was measured by an rpoB PCR-based reaction. The results demonstrated that the...
Tipo: Text Palavras-chave: RpoB gene; PCR; Ionizing irradiation; Freezing; DNA extraction; Cold smoked salmon.
Ano: 2005 URL: http://archimer.ifremer.fr/doc/2005/publication-1106.pdf
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Detection of ostreid herpesvirus-1 (OsHV-1) by PCR using a rapid and simple method of DNA extraction from oyster larvae ArchiMer
Batista, Frederico; Taris, Nicolas; Boudry, Pierre; Renault, Tristan.
A DNA extraction procedure was developed for the detection of ostreid herpesvirus-1 (OsHV-1) using the polymerase chain reaction (PCR) in oyster larvae. The DNA extraction procedure developed was tested on 8 larval samples. Abnormal nuclei with characteristic features associated with OsHV-1 infections were only observed in samples in which the viral DNA was detected by PCR. A previously described competitive PCR method was applied to detect inhibition during PCR reactions. The results show that the method can be used on small amounts of oyster larvae (3 mg) for the detection of OsHV-1 DNA by PCR.
Tipo: Text Palavras-chave: DNA extraction; Detection; Larvae; Oyster; OsHV 1; Herpesvirus.
Ano: 2005 URL: http://archimer.ifremer.fr/doc/2005/publication-2916.pdf
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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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A standardized protocol for genomic DNA isolation from Terminalia arjuna for genetic diversity analysis Electron. J. Biotechnol.
Sarwat,Maryam; Singh Negi,Madan; Lakshmikumaran,Malathi; Kumar Tyagi,Akhilesh; Das,Sandip; Shankar Srivastava,Prem.
For studying genetic diversity in natural populations of Terminalia, a medicinal plant, our attempts to isolate high quality DNA using several previously reported protocols and even modifications were unsuccessful. We therefore combined CTAB based isolation, and column based purification step, to isolate DNA from Terminalia arjuna. The DNA isolated using this standardized protocol was high in quality and suitable for restriction digestion and generation of random amplification of polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP).
Tipo: Journal article Palavras-chave: DNA extraction; Medicinal plants; Molecular markers; Terminalia species.
Ano: 2006 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000100011
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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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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
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Comparison of different methodologies for DNA extraction from Aegla longirostri BABT
Bitencourt,João Vitor Trindade; Roratto,Paula Angélica; Bartholomei-Santos,Marlise Ladvocat; Santos,Sandro.
The aim of this study was to compare some DNA extraction methodologies for Aegla longirostri. The protocols were based on the traditional phenol-chloroform DNA extraction methodology and using a commercial kit for DNA extraction. They differed in tissues used, the addition - or not - of beta-mercaptoethanol to the lysis buffer, times and methods for the animal's conservation (frozen, in ethanol or fresh). Individuals stored at -20°C for a long time supplied lower molecular weight DNA than those stored for a short time. The best yield for the specimens preserved in ethanol was obtained for 15 days storage in 95% ethanol. The kit resulted in a low quantity of high molecular weight DNA. The best protocol for DNA extraction from Aeglidae, and probably for...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Anomura; Aeglidae; Freshwater crab; DNA extraction.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132007000700010
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A simple method for isolation of genomic DNA from fresh and dry leaves of Terminalia arjuna (Roxb.) Wight and Arnot Electron. J. Biotechnol.
Deshmukh,Vishal P; Thakare,Prashant V; Chaudhari,Uddhav S; Gawande,Prashant A.
Current protocols for isolation of genomic DNA from Terminalia arjuna have their own limitations due to the presence of high content of gummy polysaccharides and polyphenols. DNA isolated by these protocols is contaminated with a yellowish, sticky and viscous matrix. In our modified DNA isolation method polysaccharides and polyphenols are removed prior to the precipitation of the DNA. Then the genomic DNA was precipitated using isopropanol. This protocol yielded a high molecular weight DNA isolated from fresh as well as dry leaves of T. arjuna, which was free from contamination and colour. Isolated DNA can be used for restriction digestion and PCR amplification. The main objective of the present protocol is to provide a simple method of isolation of DNA,...
Tipo: Journal article Palavras-chave: Ayurveda; DNA extraction; Medicinal plant; Terminalia species.
Ano: 2007 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000300014
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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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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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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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An efficient and rapid DNA minipreparation procedure suitable for PCR/SSR and RAPD analyses in tropical forest tree species BABT
Alzate-Marin,Ana Lilia; Guidugli,Marcela Corbo; Soriani,Hilda Hildebrand; Martinez,Carlos Alberto; Mestriner,Moacyr Antônio.
An efficient and rapid DNA minipreparation modified method for frozen samples was developed for five tropical tree species: Copaifera langsdorffii, Hymenaea courbaril, Eugenia uniflora, Tabebuia roseo alba and Cariniana estrellensis. This procedure that dispenses the use of liquid nitrogen, phenol and the addition of proteinase K, is an adaptation of the CTAB-based DNA extraction method. The modifications included the use of PVP to eliminate the polyphenols, only one chloroform-isoamyl alcohol step and the addition of RNase immediately after extraction with chloroform. The yields of the DNA samples ranged from 25.7 to 42.1 µg from 100 mg leaf tissue. The DNA samples extracted by this method were successfully used for PCR (SSR and RAPD) analyses in these...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Copaifera langsdorffii; Hymenaea courbaril; Eugenia uniflora; Tabebuia roseo alba; Cariniana estrellensis; DNA extraction.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000500020
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Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 BJM
Matrone,M.; Keid,L.B.; Rocha,V.C.M.; Vejarano,M.P.; Ikuta,C.Y.; Rodriguez,C.A.R.; Ferreira,F.; Dias,R.A.; Ferreira Neto,J.S.
The objective of the present study was to improve the detection of B. abortus by PCR in organs of aborted fetuses from infected cows, an important mechanism to find infected herds on the eradication phase of the program. So, different DNA extraction protocols were compared, focusing the PCR detection of B. abortus in clinical samples collected from aborted fetuses or calves born from cows challenged with the 2308 B. abortus strain. Therefore, two gold standard groups were built based on classical bacteriology, formed from: 32 lungs (17 positives), 26 spleens (11 positives), 23 livers (8 positives) and 22 bronchial lymph nodes (7 positives). All samples were submitted to three DNA extraction protocols, followed by the same amplification process with the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovine; Brucellosis; Abortion; PCR; DNA extraction.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822009000300010
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A rapid and cheap protocol for preparation of PCR templates in peanut Electron. J. Biotechnol.
Wang,Chuan Tang; Wang,Xiu Zhen; Tang,Yue Yi; Zhang,Jian Cheng; Yu,Shan Lin; Xu,Jian Zhi; Bao,Zhen Min.
This paper describes a simple, low cost and reliable DNA template preparation protocol for polymerase chain reaction (PCR) using immature leaves from peanut seeds or leaves from field-grown plants. The technique may find wide utility in studies involving PCR-based molecular markers, rapid screening for transformants and gene cloning.
Tipo: Journal article Palavras-chave: DNA extraction; Groundnut; PCR; Peanut.
Ano: 2009 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582009000200009
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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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