NucleoSpin® RNA
RNA isolation / purification Tissue - Human Kidney
Protocol tips
| Upstream tips |
|---|
| - Aliquot rDNase and store at -20 °C. |
| Protocol tips |
| - Dry the columns very well after the ethanol wash by adding an additional 2' >10000 rpm centrifuge with no buffer. |
Publication protocol
Real-time RT-PCR was performed as recently described (25). In brief, total RNA from snap-frozen biopsies was extracted using the NucleoSpin RNA Kit (Macherey-Nagel, Düren, Germany) according to the manufacturer's instructions. Integrity of RNA was checked using a NanoDrop™ 2000c spectrophotometer. Real-time reverse transcription polymerase chain reaction (RT-PCR) for gene expression analysis was performed with the ABI PRISM 7500 Sequence Detection System (Life Technologies, Carlsbad, CA, USA). Primers were directly purchased as Taqman® gene expression assays (Life Technologies, Carlsbad, CA, USA) (Supplemental Table 2). Specific gene expression was normalized to the housekeeping gene hypoxanthine-guanine phosphoribosyltransferase (HPRT) using the formula 2−ΔCt.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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