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

RNA was isolated using the TRIZOL isolation protocol (Invitrogen, Carlsbad, CA) with slight modifications. In the case of murine stomach tissue, the tissue was homogenized with gentleMACS™ Dissociator (Miltenyi Biotec, San Diego, CA). RNA was reverse transcribed using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA). For real-time rtPCR, we used the relative gene expression method. Glyceraldehyde 3-phosphate dehydrogenase (Gapdh) served as the normalizer, and tissue from uninfected WT mouse stomachs served as the calibrator sample. All real-time rtPCR was performed using an Applied Biosystems StepOne Plus real-time PCR instrument. Levels of gene transcription are indicated as “relative units”, based on comparison of tissue from H. pylori-infected mice with tissue from uninfected mice (calibrator tissue). In experiments on primary mouse epithelial cells and gastroids, levels of gene expression are indicated as “relative units”, based on comparison of stimulated cultures with unstimulated cultures (calibrator). Transcription of DefB3, Lcn2 and S100a9 were not always detected in the unstimulated cultures. When that was the case, the relative unit value was derived by assigning a CT of 40 to the unstimulated sample. Primer and probe sets were purchased as Taqman Gene Expression Assays from Applied Biosystems

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Tissue - Mouse Stomach using TRIzol Reagent from Thermo Fisher Scientific.

Manufacturer protocol

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

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Tissue - Mouse Stomach using TRIzol Reagent from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.