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Using silica particles to isolate total RNA from plant tissues recalcitrant to extraction in guanidine thiocyanate.

Analytical biochemistry (2008-01-02)
Ling-Wen Ding, Qiao-Yang Sun, Zhao-Yu Wang, Yong-Bin Sun, Zeng-Fu Xu
RESUMEN

The most commonly used protocol of the RNA isolation, the guanidine thiocyanate method, was unsuitable for recalcitrant plant tissues containing a large amount of storage proteins and secondary metabolites. We demonstrated that RNA could bind to the silica particles, which have been used successfully in DNA isolation from various sources, under a high concentration of NaCl in the presence of ethanol and sodium acetate. Based on this observation, an efficient, inexpensive, and highly reproducible technique, the acid phenol-silica method, was developed to isolate high-quality RNAs from various plant tissues recalcitrant to extraction in guanidine thiocyanate.

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Sigma-Aldrich
Guanidine thiocyanate solution, BioUltra, for molecular biology, ~6 M in H2O
Sigma-Aldrich
Tiocianato de guanidina, BioReagent, for molecular biology, ≥99%
Sigma-Aldrich
Tiocianato de guanidina, for molecular biology, ≥99%
Sigma-Aldrich
Tiocianato de guanidina, BioUltra, for molecular biology, ≥99.0% (AT)
Sigma-Aldrich
Tiocianato de guanidina, ≥97% (titration)