RNA isolation / purification Bacteria - Gram negative
Generally, isolating RNA from Gram-negative bacteria is easy. However keeping your working environment clean and RNase free (use RNase inhibitor) is essential. Some common points to keep in mind: a) Use fresh samples for isolation or store them by freezing in RNA stabilizing buffer until use. b) Choose the bacterial input amounts carefully, to ensure buffer volumes are adequate and not to overload the columns.
Start discussionFound 1 discussion for this experiment
Discussion
4 years ago
4 years ago by
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?
No matching solutions found
We'd love it if you would contribute to the research community by telling us about the product you've used to perform this experiment!
Outsource your experiment
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment