TRI Reagent™ Solution
RNA isolation / purification Bacteria - Gram negative Salmonella typhi
Publication protocol
Inner and outer foreskin pieces and colon biopsies were preserved in RNAlater (Life Technologies) and stored at −80°C. RNA isolations were carried out by placing the thawed tissue in 1ml of TRI Reagent (Life Technologies) and homogenizing it with a homogenizer with wide probe (Omni). Nucleic acid separation used 1-bromo-3-chloropropane (Sigma-Aldrich) and Ribopure Kit (Life Technologies) columns. Blood was collected in Tempus tubes, with RNA isolated according to the Tempus Spin RNA Isolation Kit (Life Technologies).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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