RNeasy Plus Mini Kit

RNA isolation / purification Bacteria - Gram negative Escherichia coli

Experiment
RNA isolation / purification Bacteria - Gram negative Escherichia coli
Product
RNeasy Plus Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Downstream tips
Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60C.

Use water to elute the RNA that is warmed to ~60C.

Publication protocol

Bacterial cells were harvested from 2 ml cultures at the logarithmic phase (OD600≈0.8). The cultures were grown in Dulbecco’s Modified Eagle Medium (DMEM) medium (Gibco) or tryptone water containing 1.5% Bacto Tryptone (Difco) and 0.5% sodium chloride at 37°C. Total RNA was extracted using the RNAprotect Bacteria Reagent and RNeasy Plus Mini Kit (Qiagen) according to the manufacturer’s instructions. The RNA samples were treated with DNase and purified using the RNeasy MiniElute Cleanup kit (Qiagen). RT-PCR reactions were carried out using 1 µg of purified RNA and the SuperScript II One Step RT-PCR system with Platinum Taq High Fidelity (Invitrogen) following the manufacturer’s instructions.

Full paper   Login or join for free to view the full paper.

Reviews

RNeasy Plus Mini Kit from Qiagen has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

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?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Bacteria - Gram negative Escherichia coli using RNeasy Plus Mini Kit from Qiagen.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Bacteria - Gram negative Escherichia coli using RNeasy Plus Mini Kit from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms