TRIzol Reagent

RNA isolation / purification Bacteria - Gram positive Clostridum botulinum

Experiment
RNA isolation / purification Bacteria - Gram positive Clostridum botulinum
Product
TRIzol Reagent from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Publication protocol

Total RNA was extracted from BoNT/A-treated cells using TRIzol reagent (Invitrogen, Carlsbad, CA) and purified using RNeasy columns (Qiagen, Valencia, CA) according to the manufacturers’ protocols. After contaminating DNA was degraded with 20 U RNase-free DNase (Promega), the concentration and purity of RNA samples were evaluated with denaturing gel electrophoresis and by measuring the absorbance at 260 and 280 nm with an Agilent 2100 analyzer (Agilent Technologies, Palo Alto, CA). RNA preparations for microarray were independently performed in triplicate under the same conditions.

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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 positive Clostridum botulinum using TRIzol Reagent from Thermo Fisher Scientific.

Paper title
Botulinum Neurotoxin Type A Induces TLR2-Mediated Inflammatory Responses in Macrophages
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for TRIzol Reagent below.

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Videos

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