RiboMinus™ Transcriptome Isolation Kit, bacteria

RNA isolation / purification Bacteria - Gram negative Pseudomonas aeruginosa

Experiment

RNA isolation / purification Bacteria - Gram negative Pseudomonas aeruginosa

Product

RiboMinus™ Transcriptome Isolation Kit, bacteria from Thermo Fisher Scientific

Manufacturer

Thermo Fisher Scientific

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Protocol tips

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	The total RNA was isolated using the illustra RNAspin kit™ (Ge Healthcare, UK) and ribosomal RNA depletion was performed using the RiboMinus™ transcriptome Kits (TermoFisher Scientific), followed the manufacturer's recommendation. The integrity of RNA was checked in agarose gel (1.5%) in a denaturant condition and the amount of RNA was determined using a spectrophotometer (Figures S7, S8) (BioTek Sinergy HT).

Protocol tips
The total RNA was isolated using the illustra RNAspin kit™ (Ge Healthcare, UK) and ribosomal RNA depletion was performed using the RiboMinus™ transcriptome Kits (TermoFisher Scientific), followed the manufacturer's recommendation. The integrity of RNA was checked in agarose gel (1.5%) in a denaturant condition and the amount of RNA was determined using a spectrophotometer (Figures S7, S8) (BioTek Sinergy HT).

Protocol tips

The total RNA was isolated using the illustra RNAspin kit™ (Ge Healthcare, UK) and ribosomal RNA depletion was performed using the RiboMinus™ transcriptome Kits (TermoFisher Scientific), followed the manufacturer's recommendation. The integrity of RNA was checked in agarose gel (1.5%) in a denaturant condition and the amount of RNA was determined using a spectrophotometer (Figures S7, S8) (BioTek Sinergy HT).

Publication protocol

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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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Manufacturer protocol

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