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 each cell line using Trizolc according to the manufacturer’s protocol and quantified using a biophotometer.d RNA was cleaned up using the RNeasy Mini kite according to the manufacturer’s protocol. RNA was reverse transcribed using Superscript II reverse transcriptasec according to the manufacturer’s protocol. PCR was carried out with the SYBR Green PCR Master Mix from PE Applied Biosystems on an ABI Prism 7700 Sequence Detection Systemf using 0.5 Amol/L of each primer and 2 μL AL cDNA template in each 25 μL AL reaction. TATA binding protein (TBP) was used as an exogenous control to compare the relative amount of target gene in different samples.

The PCR profile was as follows: 1 cycle of 95°C for 10 minutes, then 40 cycles of 95°C for 15 seconds and 60°C for 1 minute. The comparative CT method was used for relative quantitation of samples. Primers were purchased from Integrated DNA Technologies

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Manufacturer protocol

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

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