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

Small intestinal biopsies were stored in RNA stabilization solution RNAlater® (Ambion, Life Technologies, Carlsbad, CA) post assay and for total RNA purification lysed in 400 μl of TRIzol (Invitrogen, Life Technologies, Carlsbad, CA). Chloroform (80 μl) was added and whirl mixed for 15 sec. Homogenates were incubated 2–3 min at RT and centrifuged for 15 min at 10,000g (4°C). The water phase containing the RNA was further purified using RNeasy MinElute Cleanup Kit (Qiagen, Venlo, Netherlands). RNA concentration measured on a nano-drop 2000 (Thermo Scientific) and the integrity was confirmed on an Agilent 2100 Bioanalyzer using total RNA nano chips (Agilent Technologies, Santa Clara, CA, USA). RNA analysis of explant biopsies from 8 CD patients were performed, grading the quality post assay to prove cell survival. The RNA Integrity Number (RIN) score of 0 through 10 (perfect quality) is the gold standard for evaluating the integrity of the RNA

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Manufacturer protocol

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

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