TRIzol Reagent

RNA isolation / purification Tissue - Mouse Vagina

Experiment
RNA isolation / purification Tissue - Mouse Vagina
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

Isolated FRT tissues were chopped into small pieces, collected in TRIzol tubes filled with 3 mm of zirconium beads, and homogenized in three 10-s pulses using a Beadbug microtube homogenizer (Benchmark Scientific). RNA was isolated with TRIzol reagent according to the manufacturer’s instructions (Invitrogen). 1–5 µg RNA was reverse transcribed into cDNA with a Maxima First Strand cDNA Synthesis kit with dsDNase (Thermo Fisher Scientific). Real-time PCR was performed using 2x SensiFAST probe Hi-Rox mix (Bioline) with gene-specific primers (Table S1) and run on an ABIPrism 7900 sequence detector with the ΔΔCt method from SDS 2.4 software (Applied Biosystems). The relative expression of genes was calculated with the formula 2−ΔCt, where ΔCt = Ct target gene − Ct endogenous control gene. Gapdh was used as the endogenous control housekeeping gene. Viral burden was determined from a standard curve produced using serial 10-fold dilutions of ZIKV RNA or LCMV plasmid.

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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 Vagina using TRIzol Reagent from Thermo Fisher Scientific.

Paper title
Dampened antiviral immunity to intravaginal exposure to RNA viral pathogens allows enhanced viral replication
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Manufacturer protocol

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

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