RNeasy Mini Kit

RNA isolation / purification Bacteria - Gram negative Legionella pneumophilia

Experiment

RNA isolation / purification Bacteria - Gram negative Legionella pneumophilia

Product

RNeasy Mini Kit from Qiagen

Manufacturer

Qiagen

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Tips Publication protocol Reviews Discussion Papers Manufacturer protocol Videos

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).	"Include DNAse treatment for 15-20min. Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. Use water to elute the RNA that is warmed to ~60`C"

Protocol tips
To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).
Downstream tips
"Include DNAse treatment for 15-20min. Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. Use water to elute the RNA that is warmed to ~60`C"

Publication protocol

At 17 h (post-exponential phase), a 35 µl aliquot of culture was treated with RNAprotect bacteria reagent according to the manufacturer’s instructions (Qiagen), and pellets were stored at −80 °C until total RNA isolation using the RNeasy Mini kit as per the manufacturer’s instructions (Qiagen) with the exception of eluting the RNA in diethylpyrocarbonate (DEPC)-treated water (Ambion, Life Technologies).

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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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Papers

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

Paper title

Regulatory control of temporally expressed integration host factor (IHF) in Legionella pneumophila.

Full paper

Manufacturer protocol

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

Download manufacturer protocol

Videos

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