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 extracted from each sample individually using TRIzol reagent (Invitrogen, Carlsbad, CA). First-strand cDNA was generated by using a T7-linked-(dT)24 primer. After second-strand synthesis, the double-stranded cDNA was used for in vitro transcription using the ENZO BioArray High-Yield RNA transcript labeling kit (ENZO, Farmingdale, NY). The labeled cRNA was purified and fragmented. Each fragmented, biotin-labeled cRNA sample (20 µg) was hybridized to an Affymetrix mouse 430A chip. After scanning, array images were assessed by eye to confirm scanner alignment and the absence of significant bubbles or scratches. Chips had to meet the following criteria to be included in the data analysis: the number of probe pairs called present had to be greater than or equal to 30%, all spiked-in bacterial controls had to be called present, background had to be <200, the number of outliers had to be <500, and the 3V/5V ratio of GAPDH and actin had to be <2.

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Manufacturer protocol

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

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