NucleoSpin® RNA
RNA isolation / purification Bacteria - Gram negative Bordetella pertussis
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
Total RNA was isolated using a NucleoSpin RNA kit (catalog no. 740955; Macherey-Nagel, Germany) and quantified using a NanoDrop spectrophotometer (Thermo Fisher Scientific, USA). Isolated RNA was reverse transcribed into first-strand cDNA using a high-capacity cDNA reverse transcription kit according to the manufacturer's instructions (Life Technologies, USA).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
Some help with RNA isolation using Trizol
Hello! I used Trizol to extract total RNA from in-vitro cultured bacteria (1 X 10^8 cells). After phase separation, I mixed ~0.4 ml of the upper phase which contains RNA with 0.5 mL cold isopropanol. However, the amount of RNA when measured in Nanodrop was very low. In addition, the ratio between 260 and 230 was around 0.1 to 0.5. Is there a chance that my sample was contaminated by the Trizol reagent? When I collected the aqueous phase I made sure to not touch the lower phase. What should I do?
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Manufacturer protocol
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