TRIzol Reagent
RNA isolation / purification Tissue - Mouse Bone
Protocol tips
| Protocol tips |
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| - 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).Full paper Login or join for free to view the full paper.
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Discussion
Discussion
4 years ago
Author:
Paul G. Macon
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
4 years ago
Author:
Aaron Stege
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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