Consulta: subjectFacets:"RNA isolation"
Registros recuperados: 8
Data/hora: 01/06/2024 09:20:53

Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
Provedor de dados: 69
Autores: Vasanthaiah,Hemanth K.N.; Katam,Ramesh; Sheikh,Mehboob B.
Extraction of RNA from plant tissue containing high levels of polyphenols and polysaccharides is tedious and difficult in grapes. Although several protocols have been published for plant RNA isolation, most have failed to yield high quality RNA in sufficient quantity from mature and diseased grape tissue. We describe a protocol for isolating intact and high quality RNA from various grape tissues as evident by high A260/A280 absorbance ratio (1.8 to 1.9) and electrophoretic profile on denaturing formaldehyde agarose gel. On an average, 205 µg RNA per g of fresh tissue were obtained using this modified protocol. RNA quality was further assessed through RT-PCR, differential display RT-PCR and subtractive hybridization, and found to be suitable for molecular...
Tipo: Journal article
Palavras-chave: Differential display RT-PCR; Grape; RNA isolation; RT-PCR; Subtractive hybridization; Vitis.
Ano: 2008
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000300005

Comparison of two total RNA extraction protocols using the marine gorgonian coral Pseudopterogorgia elisabethae and its symbiont Symbiodinium sp.
Provedor de dados: 69
Autores: Santiago-Vázquez,Lory Z.; Ranzer,Llanie K.; Kerr,Russell G.
Marine invertebrates such as soft corals are important sources of secondary metabolites with promising biomedical applications and commercial value. RNA isolation in conjunction with reverse-transcriptase polymerase chain reaction (RT-PCR) are valuable tools utilized to study the molecular elements involved in secondary metabolite production and functional genomics. Two total RNA extraction protocols were compared using fresh tissue and flash frozen preparations from the coral Pseudopterogorgia elisabethae and from its symbiont Symbiodinium sp. isolated using RNeasy minicolumns (&lt;a href="http://www1.qiagen.com/SelectCountry.aspx"&gt;Qiagen®&lt;/a&gt;) and Trizol reagent (&lt;a href="http://www.invitrogen.com/"&gt;Invitrogen®&lt;/a&gt;). In general,...
Tipo: Journal article
Palavras-chave: Dinoflagellate; RNA isolation; RNeasy minicolumns; Trizol reagent.
Ano: 2006
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500017

Improved isolation of good-quality total RNA from the optic stalk of Mud crab, Scylla paramamosain
Provedor de dados: 69
Autores: Zhu,Houze; Jiang,Keji; Zhang,Fengying; Zhang,Dan; Pi,Yan; Ma,Lingbo.
An improved and efficient protocol was developed based on the TaKaRa RNAiso Plus Kit (Code: D9108A) for isolating good-quality total RNA from the optic stalk of mud crab, Scylla paramamosain. The protocol was based on the Trizol method with modifications. The carapace overlapping the optic stalk was retained with RNA in regular protocol. In order to remove the abundant deposition correlative with the carapace which makes the isolation of RNA particularly difficult, 5M potassium acetate solution (pH = 6.0) was added before the precipitation of RNA, and the temperature of RNA deposition was also decreased to -70ºC to ensure the stabilization of RNA. Good-quality total RNA from the optic stalk of S. paramamosain could be easily isolated with this modified...
Tipo: Journal article
Palavras-chave: Optic stalk; Potassium acetate; RNA isolation; Scylla paramamosain.
Ano: 2012
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000200005

Isolation of high quality RNA from Polyporus umbellatus (Pers.) Fries
Provedor de dados: 69
Autores: Zhang,Yuejin; Qin,Yali; Guo,Lijun; Zhou,Zili; Liang,Zongsuo; Zhang,Chenlu; Guo,Hongbo.
Background: The dried sclerotium of medicinal fungus Polyporus umbellatus (Pers.) Fries has many pharmacological functions such as diuretic and anticancer activity, in which high-content polysaccharides may play an important role. However, RNA isolation is difficult in filamentous fungi and lacking in P. umbellatus. Results: Five methods for RNA extraction from five strains collected from four provinces were assessed for their ability to recover a high-quality RNA applicable for sequence-related amplification polymorphism (SRAP) PCR and GDP-D-mannose pyrophosphorylase (GMP) gene expression profiles. Both A260/A280 and A260/A230 ratios of the best Trizol Plus + RNAiso-mate for Plant Tissue method are around 2 with a yield of 1122.00 ± 0.21 ng &amp;#956;l-1....
Tipo: Journal article
Palavras-chave: Chuling; Medicinal fungus; Polyporus umbellatus; RNA isolation.
Ano: 2012
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500010

Isolation of functional total RNA from Argemone mexicana tissues
Provedor de dados: 69
Autores: Rubio-Piña,Jorge A; Vázquez-Flota,Felipe A.
RNA extraction from recalcitrant plant tissues is frequently complicated by the presence of secondary metabolites, polysaccharides and polyphenols. These compounds may co precipitate with RNA, often rendering it unsuitable for either cDNA synthesis or hybridization in northern blot analyses and therefore, interfering with the gene analysis expression in such tissues. We have developed an efficient RNA extraction method from A. mexicana tissues. The procedure includes the use of polyvinylpolypyrrolidone (PVPP), to remove secondary metabolites, proteins and polyphenols, and a two-step precipitation with LiCl, to eliminate polysaccharides, and thus increasing RNA yield. The quality of the resulting RNA was evaluated spectrophotometrically and by agarose gel...
Tipo: Journal article
Palavras-chave: Argemone mexicana; Plant tissues; RNA isolation; Secondary metabolites.
Ano: 2008
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000400015

Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants
Provedor de dados: 69
Autores: Rubio-Piña,Jorge A; Zapata-Pérez,Omar.
RNA extraction from mangrove tissues can be difficult due to the presence of polyphenolic and polysaccharide compounds upon cell disruption. Besides, a successful RNA isolation from mangrove tissues sometimes can be strain and species specific. Two different methods were used to extract RNA from tissues (stems, leaves and roots) of black mangrove (Avicennia germinans) and white mangrove (Laguncularia racemosa), collected from the mangrove area at Progreso, Yucatan, Mexico. An optimized and modified total RNA isolation method was developed for these recalcitrant species. The protocol is based on the CTAB method including &amp;#946;-mercaptoethanol and PVPP with sequential Phenol/Chloroform/isoamyl alcohol extractions to remove protein, and polyphenols,...
Tipo: Journal article
Palavras-chave: Mangrove; Plant tissues; Polysaccharides; RNA isolation.
Ano: 2011
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000500011

A Rapid and Efficient Method for Purifying High Quality Total RNA from Peaches (Prunus persica) for Functional Genomics Analyses
Provedor de dados: 48
Autores: MEISEL,LEE; FONSECA,BEATRIZ; GONZÁLEZ,SUSANA; BAEZA-YATES,RICARDO; CAMBIAZO,VERONICA; CAMPOS,REINALDO; GONZALEZ,MAURICIO; ORELLANA,ARIEL; RETAMALES,JULIO; SILVA,HERMAN.
Prunus persica has been proposed as a genomic model for deciduous trees and the Rosaceae family. Optimized protocols for RNA isolation are necessary to further advance studies in this model species such that functional genomics analyses may be performed. Here we present an optimized protocol to rapidly and efficiently purify high quality total RNA from peach fruits (Prunus persica). Isolating high-quality RNA from fruit tissue is often difficult due to large quantities of polysaccharides and polyphenolic compounds that accumulate in this tissue and co-purify with the RNA. Here we demonstrate that a modified version of the method used to isolate RNA from pine trees and the woody plant Cinnamomun tenuipilum is ideal for isolating high quality RNA from the...
Tipo: Journal article
Palavras-chave: CDNA library; Fruit; Fruit trees; Functional genomics; Peach; Polysaccharides; Prunus persica; RNA isolation; Rosaceae; RT-PCR.
Ano: 2005
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602005000100010

Protocol adjustment improves the extraction of high-quality total RNA from common bean stems infected by Sclerotinia sclerotiorum
Provedor de dados: 64
Autores: Miranda,Rafael Novais de; Silva,Caroline Marcela da; Porto,Antonio Carlos da Mota; Pereira,Welison Andrade.
ABSTRACT The Straw Test is an assay developed to evaluate the resistance of common bean to white mold, in which the plant stems are inoculated and the symptoms of the disease are monitored. It is plausible to admit that investigating gene expression in pathogen-infected tissues may be strategically interesting. However, obtaining a quality RNA is a basic requirement for this purpose. Therefore, the objective of this study was to evaluate adjustments in protocols of commercial kits in the expectation of improving the quality of RNA obtained from bean stems. For this, plants of two lines were inoculated and the stems pathogen-infected were collected 72 hours after. For RNA extraction, two commercial reagents were used following the manufacturer’s...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Phaseolus vulgaris L.; RNA isolation; White mold; Plant-pathogen interaction.
Ano: 2019
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-70542019000100213