Total RNA Extraction Kit for Gram Negative Bacteria

September 07, 2018

BlueskyReddit
University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

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Company:

New England Biolabs

Product Name:

Monarch Total RNA Miniprep Kit

Catalog Number:

T2010S

I have recently used New England Biolabs Total RNA Miniprep kit to extract RNA from Gram negative bacterial cell pellets. Using 10^10 cells per column, I got around 30 microgram of high quality purified total RNA with 260:280 ratio of 2.2. The extracted RNA was used for qPCR analysis.

Experimental Design and Results Summary

Application

Total RNA extraction from Gram negative bacteria for gene expression analysis

Starting Material

10^10 cells lysed with lysosome

Protocol Overview

The protocol for total RNA extraction was very simple. First lyse the cell and then bind and purify RNA with the spin columns (silica). Follow the step by step kit's instructions for extraction. Finally elute the total RNA with 100 microliter of nuclease free molecular biology grade water. Check the quality of RNA and quantity using Nanodrop spectrophotometer.

Tips

For high yield, lyse the bacterial cell pellets thoroughly with lysosome (gram negative bacteria).

Results Summary

The eluted RNA quantity was 30 microgram per 10^10 cells used. The quality was checked by Nanodrop spectrometer and 260:28 ratio was 2.2 (which is of high quality). Also performed Agilent Bioanalyser analysis using Agilent RNA6000 Nano kit and found RIN values of 9.5.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

High quality RNA extraction as evidenced by Nanodrop and Agilent Bioanalyser analysis. You can also extract small RNAs using this kit.

The Bad

Nothing

The Bottom Line

Easy to use and simple protocol for Total RNA extraction. Satisfied using this kit and recommend this kit for high quality RNA extraction intended for gene expression analysis using NGS platform.

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