TRIzol Reagent

RNA isolation / purification Tissue - Mouse Stomach

Experiment
RNA isolation / purification Tissue - Mouse Stomach
Product
TRIzol Reagent from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

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

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Discussion

Discussion

2 years ago

Author: Paul G. Macon United States

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Discussion

3 years ago

Author: Aaron Stege Netherlands

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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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.

Paper title
IL-17a and IL-22 Induce Expression of Antimicrobials in Gastrointestinal Epithelial Cells and May Contribute to Epithelial Cell Defense against
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Manufacturer protocol

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

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Videos

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