RNeasy Lipid Tissue Mini Kit
RNA isolation / purification Tissue - Mouse Liver
Publication protocol
RNA extraction using currently available methods: We initially attempted to extract the RNA from in vitro cell culture and LT samples of different animal species using TRIzol (Ambion® Life Technologies™, Foster City, CA), commercially available kits, such as Exiqon’s miRCURY™ RNA Isolation Kits (Exiqon, Vedbaek, Denmark), Qiagen AllPrep DNA/RNA/Protein Mini Kit (Qiagen, Doncaster, VIC, Australia), Qiagen RNeasy Lipid Tissue Mini Kit (Qiagen), and Qiagen RNeasy Mini Kit (Qiagen) and other commonly used methods, like sodium dodecyl sulfate (SDS)‐phenol. For all these methods, LT sample (5 mg) was grounded into a fine powder in liquid nitrogen with prechilled mortar and pestle, mixed immediately with TRIzol (Ambion® Life technologiesTM, Foster City, CA)/lysis buffer provided with kits and the extraction was proceeded following the manufacturer’s instructions. Subsequently, we tried the RNA extraction after introducing the following modifications in these existing approaches: (a) SDS‐phenol extraction followed by TRIzol® (Ambion® Life Technologies™, Foster City, CA) extraction; (b) TRIzol extraction followed by a column‐based extraction for the upper phase using Exiqon’s miRCURY™ RNA Isolation Kits, Qiagen AllPrep DNA/RNA/Protein Mini Kit, Qiagen RNeasy Lipid Tissue Mini Kit, and Qiagen RNeasy Mini Kit; (c) simple addition of lithium chloride (LiCl) (500 mM) and SDS (2%) in the first homogenization step of TRIzol extraction.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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Manufacturer protocol
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