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A micromethod for high throughput RNA extraction in forest trees Biol. Res.
LE PROVOST,GREGOIRE; HERRERA,RAÚL; PAIVA,JORGE AP; CHAUMEIL,PHILIPPE; SALIN,FRANCK; PLOMION,CHRISTOPHE.
A large quantity of high quality RNA is often required in the analysis of gene expression. However, RNA extraction from samples taken from woody plants is generally complex, and represents the main limitation to study gene expression, particularly in refractory species like conifers. Standard RNA extraction protocols are available but they are highly time consuming, and not adapted to large scale extraction. Here we present a high-throughput RNA extraction protocol. This protocol was adapted to a micro-scale by modifying the classical cetyltrimethylammonium (CTAB) protocol developed for pine: (i) quantity of material used (100-200 mg of sample), (ii) disruption of samples in microtube using a mechanical tissue disrupter, and (iii) the use of SSTE buffer....
Tipo: Journal article Palavras-chave: RNA extraction; Micromethod; Forest trees.
Ano: 2007 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000400003
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A Modified protocol for total RNA isolation from forage peanuts. Repositório Alice
OLIVEIRA, J. C. de; SILVA, C. C. da; SOUZA, A. P. de; CAMPOS, T. de.
The development of specific protocols for extraction of high-quality RNA are key advances in molecular techniques for elucidating biological processes and mechanisms. The forage peanut has significant economic value for use in mixed pastures. However, it is difficult to extract high-quality RNA from forage peanuts due to high contents of tannins, polysaccharides, and other secondary metabolites. Here, it is described an efficient method for obtaining high-quality, high-yield total RNA that is usable in downstream transcriptome analysis. This modifications helped eliminate the problems associated with the presence of high concentrations of secondary metabolites in forage peanut leaves, and lacked signs of degradation. O desenvolvimento de protocolos...
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: Amendoim forrageiro; Forage peanut; Cacahuetes forrajeros; Ácido ribonucleico; Metabólico secundário; Secondary metabolic; High quality; Alta calidad; RNA extraction; Extracción de ARN; Leguminosa Forrageira; Melhoramento Genético Vegetal; RNA; Extração; Rendimento; Qualidade; Forage legumes; Arachis pintoi; Plant breeding.
Ano: 2020 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1125058
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An optmized protocol for simultaneous extraction of DNA and RNA from soils BJM
Costa,Rodrigo; Gomes,Newton C.M.; Milling,Annett; Smalla,Kornelia.
In this work we report an optimized protocol for simultaneous extraction of DNA and RNA from soil matrices. Treatment of soil matrices with ethanol followed by bead-beating worked as a successful strategy to lyse the cells without considerable degradation of nucleic acids, resulting in DNA and RNA of good yield and integrity. The reverse transcribed RNA could be amplified with primers targeting a glutamine synthetase (glnA) gene fragment. From both DNA and cDNA, 16S rDNA fragments were amplified and analyzed by Denaturing Gradient Gel Electrophoresis (DGGE). The method was applied to soil and rhizosphere (strawberry and oilseed rape) samples. Two other protocols for the extraction of nucleic acids from soil were applied to the same set of samples in order...
Tipo: Info:eu-repo/semantics/article Palavras-chave: RNA extraction; Nucleic acids; Rhizosphere; Microbial communities.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200011
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COMPARISON OF RNA EXTRACTION METHODS FOR Passiflora edulis SIMS LEAVES Rev. Bras. Frutic.
LUZ,ANNY CAROLYNE DA; PRETTI,IRANY RODRIGUES; BATITUCCI,MARIA DO CARMO PIMENTEL.
ABSTRACT Functional genomic analyses require intact RNA; however, Passiflora edulis leaves are rich in secondary metabolites that interfere with RNA extraction primarily by promoting oxidative processes and by precipitating with nucleic acids. This study aimed to analyse three RNA extraction methods, Concert™ Plant RNA Reagent (Invitrogen, Carlsbad, CA, USA), TRIzol® Reagent (Invitrogen) and TRIzol® Reagent (Invitrogen)/ice -commercial products specifically designed to extract RNA, and to determine which method is the most effective for extracting RNA from the leaves of passion fruit plants. In contrast to the RNA extracted using the other 2 methods, the RNA extracted using TRIzol® Reagent (Invitrogen) did not have acceptable A260/A280 and A260/A230...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Passiflora edulis; RNA extraction; Concert™; TRIzol®; Low temperature.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-29452016000100226
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Improvement in RNA extraction from S. cerevisie by optimization in the autolysis and NH3 hydrolysis BABT
Oliveira,Antonio Martins; Oliva Neto,Pedro de.
The optimization of autolysis of Saccharomyces cerevisiae from brewery was studied aiming at the maximum ribonucleic acid extraction and yeast extract production. The best conditions for yeast autolysis was 55.2ºC, pH= 5.1 and 9.8% NaCl for 24h of processing, without the NH3 use. In these conditions, the RNA yield was 89.7%, resulting in 51.3% of dehydrated yeast extract with 57.9% protein. The use of 12.2% NH3 at 60ºC after autolysis (8h) and plasmolysis (8h) was not viable due to the reduction in the RNA yield from 89.7to78.4%. On the other hand, the thermal shock at 60ºC for 15 minutes prior to autolysis provided an increase in the yield from 89.7 to 91.4%. The autolysis, including NaCl plasmolysis in the optimized conditions was efficient, economic and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Autolysis; NaCl plasmolysis; Yeast extract; RNA extraction; Nucleotides.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132011000500019
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Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies Electron. J. Biotechnol.
Rai,Vineeta; Ghosh,Jayadri Sekhar; Dey,Nrisingha.
RNA isolation from hard and woody internodal bamboo (Bambusa balcooa) tissue is very difficult due to the presence of secondary metabolites, polysaccharides, and polyphenolics. These compounds often co-precipitate with isolated RNA and hinder downstream applications. We have developed an efficient, cost effective and reproducible RNA isolation method from hard tissue of bamboo internode. This protocol includes an additional organic solvent refinement steps to remove endogenous phenolic compounds and acidic phenol (pH 4.2) to critically stabilize RNA in extraction buffer. In addition to these, two 2M Lithium chloride washing steps were introduced to eliminate DNA and polysaccharides contamination. The RNA isolated from the present protocol was found to be...
Tipo: Journal article Palavras-chave: Acid phenol; Bambusa balcooa; Fiber specific genes; Internode; RNA extraction.
Ano: 2010 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500022
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Protocolo para extração de RNA de plantas nativas do bioma cerrado e amazônico. Infoteca-e
CORDEIRO, M. C. R.; FRAGOSO, R. da R.; SILVA, M. S.; BARROS, L. M. G..
bitstream/item/76621/1/comtec-170.pdf
Tipo: Comunicado Técnico (INFOTECA-E) Palavras-chave: Planta nativa; Extração de RNA; Native plant; RNA extraction; Indigenous organisms; Protocol.; Cerrado.; Amazonia; Plant genetics..
Ano: 2010 URL: http://www.infoteca.cnptia.embrapa.br/infoteca/handle/doc/890719
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Signal to cut-off (S/CO) ratio and detection of HCV genotype 1 by real-time PCR one-step method: is there any direct relationship? BJID
Albertoni,Guilherme; Arnoni,CP; Araújo,PRB; Carvalho,FO; Barreto,JA.
BACKGROUND: Polymerase chain reaction (PCR) methods play an essential role in providing data related to diagnosis, monitoring and treatment of hepatitis C virus (HCV) infection. EIA results are reported as ''reactive'' or ''non reactive'' and EIA S/CO ratio may also be reported as ''high'' or ''low.'' This study aimed to evaluate the performance of a real-time RT-PCR and assess whether there is relationship between S/CO and PCR results. STUDY DESIGN AND METHODS: Sera from blood donors were analyzed by Enzyme-Linked Immunosorbent Assay (ELISA) and RT-PCR assay to detect HCV infection. RESULTS: The RT-PCR assay to genotypes 1a/b showed an acceptable linear response in serial dilutions. The samples were divided into two groups based on their serological...
Tipo: Info:eu-repo/semantics/article Palavras-chave: HCV; HCV genotype 1; Real-time PCR; ELISA; RNA extraction.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702010000200006
Registros recuperados: 8
Primeira ... 1 ... Última
 

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