TRIzol Reagent

RNA isolation / purification Tissue - Mouse Bone

Experiment
RNA isolation / purification Tissue - Mouse Bone
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

For isolation of femoral cortical bone epiphyses were cut off, and bone marrow was flushed out. The hollow bone shaft was then digested twice with 0.2% collagenase (Sigma-Aldrich, Munich, Germany) in isolation buffer (70mM NaCl, 10mM NaHCO3, 60mM sorbitol, 30mM KCl, 3mM K2HPO4, 1mM CaCl2, 0.1% bovine serum albumin, 0.5% glucose, 25 mM HEPES) at 37°C for 20 minutes each under vigorous shaking. The cleared bone shaft was frozen in liquid nitrogen and then homogenized with pestle and mortar. RNA was extracted from the homogenized tissue with TRIzol reagent (Life Technologies, Darmstadt, Germany).

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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 Bone using TRIzol Reagent from Thermo Fisher Scientific.

Paper title
Impaired proteoglycan glycosylation, elevated TGF-β signaling, and abnormal osteoblast differentiation as the basis for bone fragility in a mouse model for gerodermia osteodysplastica
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Manufacturer protocol

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

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