Use of a simple method to isolate intact RNA from partially hydratedSelaginella lepidophylla plants

Valenzuela-Avendaño, Juan P.; Mota, Iván A. Estrada; Lizama-Uc, Gabriel; Perera, Ramón Souza; Valenzuela‐Soto, Elisa M.; Zúñiga-Aguilar, José Juan

doi:10.1007/bf02772713

Cited by 37 publications

(27 citation statements)

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“…The samples obtained had no absorbance at A 260/280 nm and could not be visualized. This result has been reported taking into account that the osmoprotection might be carried out by small osmolytes like glycine betaine or the oligosaccharide; it is possible that the main problems associated with RNA extraction might be caused by oligosaccharides or even monosaccharides (Fu et al 2004;Valenzuela-Avendaño et al 2005).…”

Section: Resultsmentioning

confidence: 65%

Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants

Rubio-Piña¹,

Zapata-Pérez²

2011

Electro Journal of Biotech

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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 β-mercaptoethanol and PVPP with sequential Phenol/Chloroform/isoamyl alcohol extractions to remove protein, and polyphenols, followed by two selective purifications with LiCl and sodium acetate to eliminate polysaccharides. Although, the introduced modifications are not new, their addition proved to be decisive for success in RNA isolation. This modified procedure resulted in high quantity and quality RNA. The RNA obtained is suitable for cDNA synthesis, RT-PCR experiments and the phytochelatin synthase 1 gene amplification.

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Section: Resultsmentioning

confidence: 65%

Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants

Rubio-Piña¹,

Zapata-Pérez²

2011

Electro Journal of Biotech

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“…Though numbers of reports on the RNA isolation method from different plant tissues [6,7,11,17,22] are available in literature, no one was found to be efficient enough for fieldgrown jute tissues including seedlings rich in mucilage. Khan et al [12] had formulated a method with modification in phenol chloroform extractions and precipitation reactions.…”

Section: Introductionmentioning

confidence: 99%

Isolation of RNA from Field-Grown Jute (Corchorus capsularis) Plant in Different Developmental Stages for Effective Downstream Molecular Analysis

et al. 2011

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Jute (Corchorus capsularis), as a natural fibre producing plant species, ranks next to cotton only. Today, biotechnological approach has been considered as most accepted means for any genetic improvement of plant species. However, genetic control of the fibre development in jute has not yet been explored sufficiently for desired genetic improvement. One of the major impediments in exploring the genetic architecture in this crop at molecular level is the availability of good quality RNA from field-grown plant tissues mostly due to the presence of high amount of mucilage and phenolics. Development of a suitable RNA isolation method is becoming essential for deciphering developmental stage-specific gene expression pattern related to fibre formation in this crop species. A combination of modified hot borate buffer followed by isopycnic centrifugation (termed as HBIC) was adopted and found to be the best isolation method yielding sufficient quantity (~350-500 μg/gm fresh tissue) and good quality (A(260/280) ratio 1.88 to 1.91) RNA depending on the developmental stage of stem tissue from field-grown jute plant. The poly A(+) RNA purified from total RNA isolated by the present method was found amenable to efficient RT-PCR and cDNA library construction. The present development of RNA isolation was found to be appropriate for gene expression analysis related to fibre formation in this economically important jute plant in near future.

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“…Moreover, the method can also be applied to A. mexicana cell cultures, which also are rich in phenols and sugars (Chang et al 2003). This method was developed after unsuccessful attempts with other RNA isolation protocols for polysaccharide-rich tissues (Valenzuela-Avendaño et al 2005). We noticed that the resulted after tissue grinding in the presence of buffer, rapidly turned brown perhaps, as a consequence of polysaccharide binding oxidized phenol.…”

Section: Resultsmentioning

confidence: 99%

“…However the efficiency of such methods varies depending on the composition of the employed tissues and had been inadequate in reducing polysaccharides contamination. The addition of acidic guanidinium thiocyanate or TRIzol™ (In Vitro Life Technologies, Carlsbad CA) is widely employed to inhibit RNAse activity (Chomczynski and Sacchi, 1987;Valenzuela-Avendaño et al 2005). Organic solvents like phenol and chloroform are utilized to separate RNA from proteins into two different phases.…”

mentioning

confidence: 99%

Isolation of functional total RNA from Argemone mexicana tissues

Rubio-Piña¹,

Vázquez‐Flota²

2008

Electron. J. Biotechnol.

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Use of a simple method to isolate intact RNA from partially hydratedSelaginella lepidophylla plants

Cited by 37 publications

References 0 publications

Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants

Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants

Isolation of RNA from Field-Grown Jute (Corchorus capsularis) Plant in Different Developmental Stages for Effective Downstream Molecular Analysis

Isolation of functional total RNA from Argemone mexicana tissues

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