Figure 1From: Protocol: A simple phenol-based method for 96-well extraction of high quality RNA from ArabidopsisTRIZOL 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).Back to article page