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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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A simple ethanol wash of the tissue homogenates recovers high-quality genomic DNA from Corchorus species characterized by highly acidic and proteinaceous mucilages Electron. J. Biotechnol.
Kundu,Avijit; Sarkar,Debabrata; Bhattacharjee,Amit; Topdar,Niladri; Sinha,Mohit Kumar; Mahapatra,Bikash Sinha.
A simple miniprep based on early elimination of highly acidic and proteinaceous mucilages through ethanol washing of the tissue homogenates has been developed for the extraction of genomic DNA from mature leaves and seeds of Corchorus spp. As compared to high cetyltrimethylammonium bromide (CTAB)-NaCl DNA extraction followed by ethanol-based removal of remnant mucilages from the DNA pellet, this simple miniprep consistently and reproducibly recovers high amounts of DNA with good spectral qualities at A260/A280 and A260/A230. The purified DNA is efficiently digested by restriction endonucleases, and is suitable for PCR amplification of nuclear microsatellites with expected allele sizes.
Tipo: Journal article Palavras-chave: Corchorus; DNA extraction; Microsatellite; Mucilage; PCR; Restriction digestion.
Ano: 2011 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000100010
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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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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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DNA from tissues of young mice is optimal for genotyping Electron. J. Biotechnol.
Picazo,María G; García-Olmo,Dolores C.
Background Genotyping of mice is a common procedure in animal facilities. The aim of this study was to compare the quantity and quality of DNA extracted from samples obtained from young mice (YM; 10 d old) and adult mice (AM; 12 weeks old). We collected samples from the tail and ear of YM and AM. We also sampled blood, check cells (via buccal swabs), hair and fecal pellets of AM, and biopsied distal phalanx of YM. We isolated DNA using commercial kits and determined concentrations and purity by spectrophotometry. The integrity of DNA was evaluated by agarose-gel electrophoresis and polymerase chain reaction (PCR). Results DNA in all samples was amplified successfully but the intensities of bands after electrophoresis was heterogeneous. In general, tissues...
Tipo: Journal article Palavras-chave: DNA extraction; Genotyping; Mouse.
Ano: 2015 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000200004
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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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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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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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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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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; Bovine diseases.; Molecular epidemiology..
Ano: 2006 URL: http://www.infoteca.cnptia.embrapa.br/infoteca/handle/doc/710703
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DNA extractions from deep subseafloor sediments: Novel cryogenic-mill-based procedure and comparison to existing protocols ArchiMer
Alain, Karine; Callac, Nolwenn; Ciobanu, Maria Cristina; Reynaud, Yann; Duthoit, Frederique; Jebbar, Mohamed.
Extracting DNA from deep subsurface sediments is challenging given the complexity of sediments types, low bio-masses, resting structures (spores, cysts) frequently encountered in deep sediments, and the potential presence of enzymatic inhibitors. Promising results for cell lysis efficiency were recently obtained by use of a cryogenic mill (Lipp et al., 2008). These findings encouraged us to devise a DNA extraction protocol using this tool. Thirteen procedures involving a combination of grinding in liquid nitrogen (for various durations and beating rates) with different chemical solutions (phenol, chloroform, SDS, sarkosyl, proteinase, GTC), or with use of DNA recovery kits (MagExtractor (R)) were compared. Effective DNA extraction was evaluated in terms of...
Tipo: Text Palavras-chave: Sediment; DNA extraction; Deep subsurface biosphere.
Ano: 2011 URL: http://archimer.ifremer.fr/doc/00056/16691/14364.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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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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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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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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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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Miniprep for chloroplast and mitochondria enrichment and DNA extraction Agrociencia
Morales-Gutiérrez,Ma. Emma; Gutiérrez-Espinosa,Ma. Alejandra; Santacruz-Varela,Amalio; Ramírez-Ramírez,Iván; Pérez-Grajales,Mario; González-Hernández,Víctor A..
Abstract Published protocols for chloroplast and mitochondrial DNA enrichment are based on large samples (more than 5 g) which imply high costs and plenty of sample material. Reduction of tissue sample and reagent quantities is possible and convenient. This work describes an efficient miniprep for chloroplast and mitochondria DNA enrichment and extraction from 500 mg of pericarp samples of Manzano chili fruits (Capsicum pubescens R. & P.). Extraction efficiency from fresh fruit samples was 20 µg DNA g-1 for choloroplasts and 33 µg DNA g-1 for mitochondria. Isolation using the recommended miniprep resulted in costs reduction of 160x for chloroplast and up to 8x for mitochondria. PCR amplification of reference marker genes for chloroplast (rbcL, Rubisco...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Capsicum pubescens; Chloroplast DNA; Mitochondria DNA; DNA extraction.
Ano: 2016 URL: http://www.scielo.org.mx/scielo.php?script=sci_arttext&pid=S1405-31952016000800977
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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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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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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
Registros recuperados: 34
Primeira ... 12 ... Última
 

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