TRIzol Reagent

RNA isolation / purification Tissue - Rat Pancreas

Experiment
RNA isolation / purification Tissue - Rat Pancreas
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

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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Discussion

Discussion

4 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

5 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 - Rat Pancreas using TRIzol Reagent from Thermo Fisher Scientific.

Paper title
Autoantigen-Induced Focusing of Vβ13 T Cells Precedes Onset of Autoimmune Diabetes in the LEW.1WR1 Rat
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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