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Figure 1 | Plant Methods

Figure 1

From: Protocol: A simple phenol-based method for 96-well extraction of high quality RNA from Arabidopsis

Figure 1

TRIZOL is not suitable for extracting RNA from very young tissues. RNA extracted using TRIZOL can satisfactorily isolate total RNA (18S bands) but very little mRNA (β-TUBULIN bands) is extracted from the very youngest Arabidopsis tissues. This bias is not evident in RNA prepared from the same tissue using P:C-L extraction. (A). Northern blot of TRIZOL and P:C-L-extracted RNA (10 μg) from samples grown for zero, three, six, or 12 days prior to a six week cold treatment and harvested in the cold. As the age of the seedlings increase, β-TUBULIN expression rises as a proportion of total RNA in the TRIZOL-extracted RNA preparations. By the time tissue was 12 days old, the bias observed in TRIZOL-extracted RNA had diminished. This effect is also observed for FLC (data not shown). (B). Northern blot comparing TRIZOL and P:C-L-extracted RNA (10 μg) of non cold treated five-day old (i.e. just germinated) Arabidopsis seedlings from several different genotypes (a-g) shows that the age-related bias observed in TRIZOL-extracted RNA is highly repeatable and not subject to growth conditions. TRIZOL and P:C-L extracted RNA preparations appear to differ significantly in the population of major RNA species visible by agarose gel electrophoresis (asterisks).

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