VWR Life Science RiboZol™ RNA Extraction Reagent
RNA isolation / purification Bacteria - Gram negative Legionella pneumophilia
Protocol tips
| Protocol tips |
|---|
| If the aqueous layer appears pink, not enough RobiZol was used |
| Downstream tips |
| For RT-PCR applications, DNase treatment might be required |
Publication protocol
Total L. pneumophila RNA was extracted from cells at an OD620 of 0.9 by the RiboZol RNA extraction reagent (Amresco, OH). Total RNA was treated with DNase I, and RNA concentrations were determined using a NanoDrop ND-1000 spectrophotometer (Nanodrop Technologies). First-strand cDNA was generated using the SuperScript first-strand synthesis system for reverse transcriptase PCR (RT-PCR) (Invitrogen, CA). When the RT reaction was completed, samples were adjusted to 200 μl with molecular biology water.Full paper Login or join for free to view the full paper.
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Discussion
Discussion
5 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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