Isolation of functional total RNA from Argemone mexicana tissues

Rubio-Piña, Jorge; Vázquez‐Flota, Felipe

doi:10.2225/vol11-issue4-fulltext-13

Cited by 10 publications

(10 citation statements)

References 6 publications

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“…They introduced the use of polyvinylpolypyrrolidone (PVPP) in the extraction buffer and LiCl precipitation. The insoluble PVPP has been used for RNA extraction from recalcitrant tissues (Rubio-Piña and Vázquez-Flota, 2008), as it binds and removes phenolic compounds. Fu et al (2004) reported that extraction buffers containing LiCl could eliminate the entangling polysaccharides from nucleic acids in Aegiceras corniculatum; however, this was not suitable for the RNA extraction of A. germinans and L. racemosa (Figure 1a).…”

Section: Resultsmentioning

confidence: 99%

“…These protocols were modified using LiCl/ sodium acetate to remove polysaccharides during the RNA precipitation steps. This approach has been used previously (Chan et al 2004;Nguyen et al 2006;Rodrigues et al 2007;Rubio-Piña and Vázquez-Flota, 2008). The results of the two RNA extraction methods used on the tissue samples are shown in Table 1.…”

Section: Resultsmentioning

confidence: 99%

“…Unfortunately, the task of extracting intact RNA is difficult due to the susceptibility of RNA molecules to the enzymatic degradation by RNase (Rubio-Piña and Vázquez-Flota, 2008). The problem of isolating RNA from mangrove is magnified by the release of high contents of polyphenolics and metabolites after disruption of the cells, which are embedded in viscous polysaccharides.…”

Section: Introductionmentioning

confidence: 99%

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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: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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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“…Total RNA was isolated as described by Rubio-Piña and Vázquez-Flota (2008) and converted to cDNA using the M-MLV reverse transcriptase (Invitrogen) in order to monitor transcript accumulation by RT-PCR. Oligonucleotide primers were designed based on the A. mexicana cDNAs for TyDC (EU.…”

Section: Gene Expression Analysismentioning

confidence: 99%

The sequential exposure to jasmonate, salicylic acid and yeast extract promotes sanguinarine accumulation in Argemone mexicana cell cultures

et al. 2011

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The effects of the sequential application of methyl jasmonate (MeJa), salicylic acid (SA) and yeast extract (YE) to Argemone mexicana cell cultures were compared to either the sole application of each elicitor, or to the three-partite mixture. The highest sanguinarine accumulation occurred using the sequential treatment (ninefold over unexposed control cultures), followed by the single application of YE (fivefold). The elicitor mixture produced less sanguinarine than sole exposure to YE but higher than MeJa alone. SA did not produce any effect. Transcripts corresponding to tyrosine decarboxylase and berberine bridge enzyme accumulated in treated cells, but did not correlate with alkaloid accumulation. Discrete epifluorescence foci, surrounding the nucleus and scattered throughout the cytoplasm of elicited cells, suggested the presence of alkaloid-accumulating vesicles which could participate in a mechanism to avoid sanguinarine toxicity.

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“…As the isolation itself is based on small dose with a few optic stalks, the common protocol with hexadecyltrimethylammonium bromide and polyvinylpolypyrrolidone (Liao et al 2004;Rubio-Piña and Vázquez-Flota, 2008;Rubio-Piña and Zapata-Pérez, 2011), or sodium dodecyl sulfate (Rai et al 2010) for plants is not very applicable to stalks as it is suitable for removing polysaccharides and polyphenols. While Trizol, a current RNA protocol will be an appropriate protocol with advantage in small dose and high quality and it is frequently used in RNA isolation (Chadderton et al 1997;Gehrig et al 2000;Bovée et al 2001).…”

Section: Introductionmentioning

confidence: 99%

Improved isolation of good-quality total RNA from the optic stalk of Mud crab, Scylla paramamosain

Zhu

Jiang

Zhang

et al. 2012

Electro Journal of Biotech

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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 protocol and three conventional methods were also employed to confirm the quality of RNA. This improved method would be helpful in facilitating molecular research of crabs involving RNA from the optic stalk.

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Isolation of functional total RNA from Argemone mexicana tissues

Cited by 10 publications

References 6 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

The sequential exposure to jasmonate, salicylic acid and yeast extract promotes sanguinarine accumulation in Argemone mexicana cell cultures

Improved isolation of good-quality total RNA from the optic stalk of Mud crab, Scylla paramamosain

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