High Pure RNA Isolation Kit

RNA isolation / purification Bacteria - Gram negative Legionella pneumophilia

Experiment
RNA isolation / purification Bacteria - Gram negative Legionella pneumophilia
Product
High Pure RNA Isolation Kit from Roche Lifesciences
Manufacturer
Roche Lifesciences

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

1 × 106 cells of A. castellanii were infected with L. pneumophila Paris at a MOI of 20 for 2 h in 6-well plates. After the infection (0 h), cells were harvested for RNA extraction at different time points 2 h, 16 h and 24 h. To increase the efficiency of the cell lysis, RNA extractions were preceded by a mechanical cell lysis using a FastPrep®-24 Instrument (MP) for 30 seconds at a speed of 5.0 (twice) with FastPrep Tubes containing glass breads 150–212 μm (Sigma). RNA extraction was performed using the High Pure RNA Isolation Kit (Roche).



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Discussion

Discussion

5 years ago

Author: Paul G. Macon United States

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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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Bacteria - Gram negative Legionella pneumophilia using High Pure RNA Isolation Kit from Roche Lifesciences.

Paper title
prevents proliferation of its natural host
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Manufacturer protocol

Download the product protocol from Roche Lifesciences for High Pure RNA Isolation Kit below.

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