NucleoSpin® RNA

RNA isolation / purification Bacteria - Gram negative Bordetella pertussis

Experiment
RNA isolation / purification Bacteria - Gram negative Bordetella pertussis
Product
NucleoSpin® RNA from Macherey Nagel
Manufacturer
Macherey Nagel

Protocol tips

Upstream tips
Aliquot rDNase and store at -20 °C.
Protocol tips
Dry the columns very well after the ethanol wash by adding an additional 2' >10000 rpm centrifuge with no buffer.

Publication protocol

Total RNA was isolated using a NucleoSpin RNA kit (catalog no. 740955; Macherey-Nagel, Germany) and quantified using a NanoDrop spectrophotometer (Thermo Fisher Scientific, USA). Isolated RNA was reverse transcribed into first-strand cDNA using a high-capacity cDNA reverse transcription kit according to the manufacturer's instructions (Life Technologies, USA).

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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 Bordetella pertussis using NucleoSpin® RNA from Macherey Nagel.

Paper title
Adenylate Cyclase Toxin Disrupts Functional Integrity of Bronchial Epithelial Layers
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Manufacturer protocol

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