Consulta: subjectFacets:"DNA extraction"
Registros recuperados: 34
Data/hora: 01/06/2024 03:32:45

A standardized protocol for genomic DNA isolation from Terminalia arjuna for genetic diversity analysis
Provedor de dados: 69
Autores: 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

A simple ethanol wash of the tissue homogenates recovers high-quality genomic DNA from Corchorus species characterized by highly acidic and proteinaceous mucilages
Provedor de dados: 69
Autores: 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

Isolation of high quality DNA: a protocol combining &amp;#8220;rennet&amp;#8221; and glass milk
Provedor de dados: 69
Autores: 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 &amp;#8220;rennet&amp;#8221; in cheese production and which is easily obtained on the commercial market. The second protocol uses &amp;#8220;rennet DNA extraction protocol&amp;#8221; combined with the DNA binding capacity of glass powder (glass milk), which can easily be &amp;#8220;home made&amp;#8221;. 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

Simple method to prepare DNA templates from a slice of peanut cotyledonary tissue for Polymerase Chain Reaction
Provedor de dados: 69
Autores: 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 &amp;#956;l. 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

DNA from tissues of young mice is optimal for genotyping
Provedor de dados: 69
Autores: 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

A simple method for isolation of genomic DNA from fresh and dry leaves of Terminalia arjuna (Roxb.) Wight and Arnot
Provedor de dados: 69
Autores: 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

A rapid and cheap protocol for preparation of PCR templates in peanut
Provedor de dados: 69
Autores: 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

Protocol for extraction of genomic DNA from swine solid tissues
Provedor de dados: 74
Autores: 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

Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
Provedor de dados: 74
Autores: 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

Extração de DNA a partir de coágulos sanguíneos bovinos.
Provedor de dados: 14
Autores: 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

DNA extractions from deep subseafloor sediments: Novel cryogenic-mill-based procedure and comparison to existing protocols
Provedor de dados: 5
Autores: 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

Detection of ostreid herpesvirus-1 (OsHV-1) by PCR using a rapid and simple method of DNA extraction from oyster larvae
Provedor de dados: 5
Autores: 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

Comparison of three methods of DNA extraction from cold-smoked salmon and impact of physical treatments
Provedor de dados: 5
Autores: 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

The use of museum specimens as source of DNA
Provedor de dados: 16
Autores: 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

การสกัดดีเอ็นเอจากข้าวโพดในระดับเล็กสำหรับทำพีซีอาร์
Provedor de dados: 106
Autores: 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

Método não-invasivo na obtenção de DNA de búfalos - DOI: 10.4025/actascianimsci.v30i4.4839
Provedor de dados: 2
Autores: 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

Miniprep for chloroplast and mitochondria enrichment and DNA extraction
Provedor de dados: 115
Autores: 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. &amp; 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

Comparison of preservation methods of Atta spp. (Hymenoptera: Formicidae) for RAPD analysis
Provedor de dados: 44
Autores: 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

Extraction of high-quality host DNA from feces and regurgitated seeds: a useful tool for vertebrate ecological studies
Provedor de dados: 48
Autores: 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

Comparison of different methodologies for DNA extraction from Aegla longirostri
Provedor de dados: 52
Autores: 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

Rapid yeast DNA extraction by boiling and freeze-thawing without using chemical reagents and DNA purification
Provedor de dados: 52
Autores: 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

Direct RAPD evaluation of bacteria without conventional DNA extraction
Provedor de dados: 52
Autores: 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

An efficient and rapid DNA minipreparation procedure suitable for PCR/SSR and RAPD analyses in tropical forest tree species
Provedor de dados: 52
Autores: 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

An Alternative and Rapid Method for the Extraction of Nucleic Acids from Ixodid Ticks by Potassium Acetate Procedure
Provedor de dados: 52
Autores: Rodríguez,Islay; Fraga,Jorge; Noda,Angel Alberto; Mayet,Milagro; Duarte,Yanisia; Echevarria,Eduardo; Fernández,Carmen.
Four variants of the potassium acetate procedure for DNA extraction from ixodid ticks at different stage of their life cycles were evaluated and compared with phenol-chloroform and ammonium hydroxide methods. The most rapid and most efficient variant was validated in the DNA extraction procedure from the engorged ticks collected from bovine, canine as well as from house ticks for the screening of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp. The ammonium hydroxide procedure was used for non-engorged ticks. All the variants were efficient and allowed obtaining PCR-quality material according to the specific amplification of 16S rRNA gene fragment of the original tick. DNA extracted from the ticks under the study was tested by multiplex PCR...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Ticks; PCR; DNA extraction.
Ano: 2014
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132014000400542

Development of an Extraction Protocol for Fetal Sexing from the Fourth Week of Gestation by Real Time PCR
Provedor de dados: 52
Autores: Oliveira,Edwin José Torres de; Rabacow,Ana Paula Maluf; Figueiredo,Bruno Pereira de; Ishikawa,Raíssa Borges; Pessatto,Lucas Roberto; Pelizaro,Bruno Ivo; Figueiredo,Helder Pereira de; Oliveira,Rodrigo Juliano.
Abstract Low number of fetal cells in maternal blood limited the use of fetal materials in diagnostic and clinical applications. This research developed a technology which allowed the extraction of fetal DNA by a non-invasive method that offers no risk to the mother or fetus. A total of 132 pregnant women participated in this inquiry. The DNA extraction was performed employing an in-house method based on guanidine thiocyanate and magnetic bead. For the amplification it was used the Quantifier Y Kit™. The fetal sexing analysis of the 132 pregnant women were 100% in agreement with the ultrasound. Sensitivity and specificity for detection of Y chromosome sequences was possible using Real-Time Polymerase Chain Reaction since the 4th week of gestation. This...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Fetal-sexing; DNA extraction; RtPCR.
Ano: 2019
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132019000100416

Urine as a promising sample for Leishmania DNA extraction in the diagnosis of visceral leishmaniasis - a review
Provedor de dados: 55
Autores: 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

Use of a cysteine proteinase from Carica candamarcensis as a protective agent during DNA extraction
Provedor de dados: 56
Autores: 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

Enhancement of soil DNA extraction by the use of a hand held mixer
Provedor de dados: 58
Autores: 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

Comparison of different protocols for the extraction of microbial DNA from reef corals
Provedor de dados: 58
Autores: Santos,H.F.; Carmo,F.L.; Leite,D.C.A.; Jesus,H.E.; De Carvalho Maalouf,P.; Almeida,C.; Soriano,A.U.; Altomari,D.; Suhett,L.; Vólaro,V.; Valoni,E.; Francisco,M.; Vieira,J.; Rocha,R.; Sardinha,B.L.; Mendes,L.B.; João,R.R.; Lacava,B.; Jesus,R.F.; Sebastian,G.V.; Pessoa,A.; van Elsas,J.D.; Rezende,R.P.; Pires,D.O.; Duarte,G.; Castro,C.B.; Rosado,A.S.; Peixoto,R.S.
This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Coral; Microbial diversity; DNA extraction.
Ano: 2012
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822012000200012

Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308
Provedor de dados: 58
Autores: 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

Assessment of the quality of dna extracted by two techniques from Mycobacterium tuberculosis for fast molecular identification and genotyping
Provedor de dados: 58
Autores: Miyata,Marcelo; Santos,Adolfo Carlos Barreto; Mendes,Natália Helena; Cunha,Eunice Atsuko; Melo,Fernando Augusto Fiúza de; Leite,Clarice Queico Fujimura.
We report a comparative study of two DNA extraction techniques, thermolysis and chemical lysis (CTAB), for molecular identification and genotyping of M. tuberculosis. Forty DNA samples were subjected to PCR and the results demonstrated that with thermolysis it is possible to obtain useful data that enables fast identification and genotyping.
Tipo: Info:eu-repo/semantics/article
Palavras-chave: DNA extraction; Mycobacterium tuberculosis; Thermolysis; CTAB.
Ano: 2011
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000200045

Comparison of DNA extraction protocols for microbial communities from soil treated with biochar
Provedor de dados: 58
Autores: Leite,D.C.A.; Balieiro,F.C.; Pires,C.A.; Madari,B.E.; Rosado,A.S.; Coutinho,H.L.C.; Peixoto,R.S.
Many studies have evaluated the effects of biochar application on soil structure and plant growth. However, there are very few studies describing the effect of biochar on native soil microbial communities. Microbial analysis of environmental samples requires accurate and reproducible methods for the extraction of DNA from samples. Because of the variety among microbial species and the strong adsorption of the phosphate backbone of the DNA molecule to biochar, extracting and purifying high quality microbial DNA from biochar-amended soil is not a trivial process and can be considerably more difficult than the extraction of DNA from other environmental samples. The aim of this study was to compare the relative efficacies of three commercial DNA extraction...
Tipo: Info:eu-repo/semantics/other
Palavras-chave: Biochar; DNA extraction; PCR-DGGE; Microbial communities; DNA purity indices.
Ano: 2014
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000100023

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
Provedor de dados: 118
Autores: 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

Comparison of nine DNA extraction methods for the diagnosis of bovine tuberculosis by real time PCR
Provedor de dados: 65
Autores: Moura,André; Hodon,Mikael Arrais; Soares Filho,Paulo Martins; Issa,Marina de Azevedo; Oliveira,Ana Paula Ferreira de; Fonseca Júnior,Antônio Augusto.
ABSTRACT: Bovine tuberculosis is an infectious disease with a high impact on the cattle industry, particularly in developing countries. PCR is a very sensitive method for detection of infectious agents, but the sensitivity of molecular diagnosis is largely dependent on the efficiency of the DNA extraction methods. The objective of this study was to evaluate DNA extraction methods for direct detection of  Mycobacterium bovis in bovine tissue. Nine commercial kits for DNA extraction were evaluated when combined with two real time PCRs. The DNeasy Blood &amp; Tissue Kit from QIAGEN showed better performance and sensitivity followed by the DNA Mini Kit RBC and FTA Elute Micro Card. Results suggested that, even when the analytical sensitivity of the qPCR is...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Bovine tuberculosis; DNA extraction; Real time PCR.
Ano: 2016
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016000701223