A modified CTAB and Trizol® protocol for high-quality RNA extraction from whole wheat seedlings, including rhizosphere

Chaparro-Encinas, L. A. and Arellano-Wattenbarger, G. L. and Parra-Cota, F. I. and de los Santos-Villalobos, S. (2020) A modified CTAB and Trizol® protocol for high-quality RNA extraction from whole wheat seedlings, including rhizosphere. CEREAL RESEARCH COMMUNICATIONS, 48 (3). pp. 275-282. ISSN 0133-3720 (print), 1788-9170 (online)

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Abstract

The RNA extraction was performed from foliar (F) and whole wheat plants (including rhizosphere) (WP) samples by (1) the standard TRIzol® protocol, and (2) a modifed CTAB and TRIzol® protocol. The modifed CTAB and TRIzol® protocol was able to extract high-quality RNA (205.96±18.7 ng/µL for F, and 231.76±66.8 ng/µL for WP; RIN>8.0), compared to the standard TRIzol® protocol (92.73±24.2 ng/µL for F, and WP completely degraded, RIN<8.0). Real-time RT-PCR assay was carried out for 6-SFT1 (target) and 18S rRNA (housekeeping) genes, which showed a PCR efciency of 111% and 118%, respectively, and a no signifcant relative expression (3773±1383.8 for F and 2847±1037.5 for WP) was observed from RNA extracted by the modifed protocol. The modifed CTAB and TRIzol® protocol was able to produce high-quality RNA (yield, purity, and integrity) from foliar and whole wheat plants (including rhizosphere with recalcitrant properties).

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