High Pure RNA Isolation Kit
RNA isolation / purification Bacteria - Gram negative Neisseria gonorrhoeae
Protocol tips
| Upstream tips |
|---|
| Be careful to create an RNase-free working environment |
| Protocol tips |
| Always mix the sample tube well after addition of each reagent. |
Publication protocol
N. gonorrhoeae drg::cm, drg::dam and FA1090 wt strains were grown on plates for 24 h as described above. Cells were then harvested from one plate and total RNA was isolated using the High Pure RNA Isolation Kit (Roche), according to the manufacturer's recommendations. Contaminating DNA were removed using DNA-free™, DNase Treatment & Removal (Ambion), according to the manufacturer's instructions. RNA concentration and RNA quality were measured using NanoDrop 2000 (Thermo Scientific) and 2100 Bioanalyzer (Agilent Technologies).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
Download the product protocol from Roche Lifesciences for High Pure RNA Isolation Kit below.
Download manufacturer protocol
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