Purification Of rRNA Depleted Total RNA Of Salmonella Enterica For RNA-Seq Library Prep

August 11, 2016

University of Birmingham
Biosciences
Research Assistant

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Quality of Results

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Company:

Qiagen

Product Name:

RNeasy MinElute Cleanup Kit (50)

Catalog Number:

74204

I used Qiagen's RNeasy MinElute Cleanup kit for purification of rRNA depleted total RNA samples from Salmonella enterica. Purification protocol was easy perform and got recovery of 80%. Quantity was 12-15 ng/microliter as determined by Qubit HS RNA assay kit method.

Experimental Design and Results Summary

Application

Purification of total RNA of Salmonella enterica after depleting rRNA

Starting Material

rRNA depleted total RNA samples

Protocol Overview

After removal/depleting rRNA from the total RNA, 80 microliter of rRNA depleted samples were made upto the volume of 100 microliter using RNase free water. Then entire 100 microliter was applied onto RNeasy MinElute columns. Lysis and washing steps were performed using lysis and washing buffer provided with kit according to kit's instructions. Finally rRNA depleted total RNA was eluted in the 12 microliter of RNase free water.

Tips

While working with RNA samples, keep samples in between on the ice. Avoid exposure of RNA samples at room temperature.

Results Summary

Recovery after purification was 80% with quantity of 12-15 ng/microliter as determined by Qubit HS RNA assay method.

Additional Notes

None

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Summary

The Good

Protocol is simple and easy to follow on. Purification of 6 samples at a time takes less than 30 minutes time.

The Bad

None

The Bottom Line

Very good and ideal kit for purification of the rRNA depleted total RNA samples especially for beginners who does not have much experience with ethanol precipitation method.

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