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 islets in TRIzol according to the manufacturers’ protocol, treated with Turbo DNA-free (Applied Biosystems, Carlsbad, CA) to prevent genomic DNA contamination and reverse transcribed into cDNA (High Capacity cDNA Reverse Transcription Kit; Applied Biosystems). Quantitative PCR was performed with an ABI 7900HT Sequence Detector using SYBR Green PCR Mix (Applied Biosystems), 0.5 μmol/L primers and 40 amplification cycles (95.0°C for 0:15; 60.0°C for 1:00). Amplification data were collected and analyzed using ABI SDS2.2 software. Fold change in transcript abundance was normalized to either glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or TCR β-chain constant region (TCR-Cβ) by the ΔΔCt method and analyzed by Student t test (two comparators) or one-way ANOVA (three comparators) to determine significant differences in transcript abundance between strains or time points

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Manufacturer protocol

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

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