RNeasy Protect Bacteria Mini Kit

RNA isolation / purification Bacteria - Gram positive Enterococcus faecalis

Experiment
RNA isolation / purification Bacteria - Gram positive Enterococcus faecalis
Product
RNeasy Protect Bacteria Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Upstream tips
To ensure reliable and reproducable gene expression, minimal media is recommended, though complex media also work with this kit.
Downstream tips
RNA protect only stabilizes the RNA and lyses the bacteria, for further purification RNeasy mini or midi kits are recommended by the manufacturer.

Publication protocol

Total RNA was isolated using the RNeasy Protect Bacteria Mini Kit (Qiagen) following the manufacturers' protocol with lysozyme lysis. The RNA was eluted in RNase free water, digested with 2 U of Turbo DNaseI (Ambion) for 30 min at 37°C, and again purified using the RNeasy Mini Kit (Qiagen). RNA was recovered in RNase free water, heat denatured for 10 min at 65°C, and quantified with the NanoDrop ND-1000 UV-Vis Spectrophotometer (NanoDrop Technologies), and a quality profile with the Agilent 2100 bioanalyzer (Agilent Technologies) was made.

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Reviews

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Discussion

Discussion

4 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 Enterococcus faecalis using RNeasy Protect Bacteria Mini Kit from Qiagen.

Paper title
The TIR Domain Containing Locus of Is Predominant among Urinary Tract Infection Isolates and Downregulates Host Inflammatory Response
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Manufacturer protocol

Download the product protocol from Qiagen for RNeasy Protect Bacteria Mini Kit below.

Download PDF Download manufacturer protocol

Videos

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