REzol C&T
RNA isolation / purification Bacteria - Gram positive Clostridium difficile
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Publication protocol
Total RNA was extracted using REzol (PROtech, Taipei, Taiwan). mRNA levels were analyzed with real-time quantitative RT-PCR (Applied Biosystems, Foster City, CA) with Actb as the reference gene in each reactionFull 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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