miRNeasy Mini kit

RNA isolation / purification Bacteria - Gram negative Bordetella pertussis

Experiment
RNA isolation / purification Bacteria - Gram negative Bordetella pertussis
Product
miRNeasy Mini kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Protocol tips
- To capture a higher amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.

Publication protocol

Total RNA was extracted from the moDCs, cultured in the presence of the indicated stimuli 6 h or 24 h post stimulation, for messenger RNA (mRNA) or microRNA (miRNA) analysis respectively. The Phenol extraction column-based miRNeasy RNA isolation kit (Qiagen, Venlo, the Netherlands) was used. Murine lung tissue collected on day 3 and day 7 after infection was homogenized by using the FastPrep®-24 bead homogenizer (MP Biomedicals, Santa Ana, CA, USA) in 700 µL Qiazol (Qiagen). RNA was isolated according to the manufacturer’s instructions with minor modifications, namely an extra column-washing step was added using 80% ethanol before extracting the RNA using RNase-free water. RNA quality and integrity was determined using Lab-on-Chip analysis on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The RNA integrity numbers (RIN) of all RNA samples were >8.



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Discussion

Discussion

2 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 Bordetella pertussis using miRNeasy Mini kit from Qiagen.

Paper title
pertactin knock-out strains reveal immunomodulatory properties of this virulence factor
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Manufacturer protocol

Download the product protocol from Qiagen for miRNeasy Mini kit below.

Download PDF Download manufacturer protocol

Videos

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