TRIzol Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Protocol tips
For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Publication protocol

Total RNA was isolated from cells using TRIzol reagent (Life Technologies #15596026) followed by chloroform extraction, isopropanol precipitation, and a 75% (v/v) ethanol wash. RNA samples (1 μg) were reverse-transcribed to cDNA using a High Capacity cDNA kit (Applied Biosystems #4368814). cDNA in 1 μl of the reaction mixture was amplified with PCR Master Mix (Life Technologies #K0171) and 10 μmol each of the sense and antisense primers. The thermal cycle profile was as follows: denaturation at 95 °C for 30 s, annealing at 55–61 °C for 30 s depending on the primers used, and extension at 72 °C for 90 s. Each PCR reaction was carried out for 35 cycles, and the PCR products were size fractionated on 1% agarose gel/GelGreen (1:10000) (Fisher Biotec #41005) and visualized under UV Trans illumination (FB Biotech).

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for TRIzol Reagent below.

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