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 (Invitrogen, Carlsbad, CA), and purified by PureLink RNA Mini Kit (Ambion, Life Technologies, Carlsbad, CA) according to the manufacturer’s instructions. cDNA was produced from 250ng of total RNA using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Life Technologies, Carlsbad CA) according to the manufacturer’s protocol. qPCR was performed using TaqMan Gene Expression Master Mix with the canine β-catenin/CTNNB1 (Cf02667771_m1) TaqMan Gene Expression Assay (Applied Biosystems) according to the manufacture’s protocol, on a Bio-Rad iQ5 Multicolor Real-Time PCR Detection System with Bio-Rad iCycler machine and iQ5 software. Ct values were normalized to 18S expression (4352930E, Applied Biosystems). Relative differences in mRNA expression of transfected/treated cells were compared to untreated cells using the ΔΔCt method.20 Gene expression of samples was measured in triplicate.

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Manufacturer protocol

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

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