TRIzol Reagent

RNA isolation / purification Tissue - Mouse Cornea

Experiment
RNA isolation / purification Tissue - Mouse Cornea
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

Real-time PCR was performed to detect gene expression in corneal epithelium and conjunctival tissues. Total RNA was extracted from mouse corneal epithelia and conjunctival tissues with the use of TRIzol (Invitrogen, Carlsbad, CA) and was reverse-transcribed to cDNA with the use of the ExScript RT Reagent kit (Takara, Dalian, China). The primers used to amplify specific gene products are listed in Table 1. Real-time PCR was performed with a StepOne Real-Time PCR detection system (Applied Biosystems, Darmstadt, Germany) with the use of a SYBR Premix Ex Taq Kit (Takara) according to the manufacturer's instructions. The amplification program included an initial denaturation step at 95°C for 10 minutes, followed by denaturation at 95°C for 10 seconds and annealing and extension at 60°C for 30 seconds for 40 cycles. The results of relative quantitative real-time PCR were analyzed by the comparative threshold cycle method and normalized to β-actin expression as an internal control.

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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 - Mouse Cornea using TRIzol Reagent from Thermo Fisher Scientific.

Paper title
Meibomian Gland Absence Related Dry Eye in Ectodysplasin A Mutant Mice.
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Manufacturer protocol

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

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