TRIzol Reagent

RNA isolation / purification Bacteria - Gram negative Vibro parahaemolyticus

Experiment
RNA isolation / purification Bacteria - Gram negative Vibro parahaemolyticus
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 RNAs were extracted using the TRIzol Reagent (Invitrogen). RNA quality was monitored by agarose gel electrophoresis, and RNA quantity was determined by spectrophotometry. The contaminated DNA in the RNA samples was removed by using the Amibion's DNA-free™ Kit. cDNAs were generated by using 5 μg of RNA and 3 μg of random hexamer primers. Real-time PCR was performed through the LightCycler system (Roche) together with the SYBR Green master mix (Zhan et al., 2008). Based on the standard curve of 16S rRNA expression for each RNA preparation, the relative mRNA levels were determined with the ΔCt values by the classic ΔCt method. 16S rRNA gene was used to normalize.

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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 Vibro parahaemolyticus using TRIzol Reagent from Thermo Fisher Scientific.

Paper title
H-NS is a repressor of major virulence gene loci in
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Manufacturer protocol

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

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