TURBO DNase Treatment To Remove DNA From RNA Preparations

February 02, 2017

Albert Einstein College of Medicine
Clinical Research
Pre-doctoral researcher

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Company:

Ambion by Life Technologies (ThermoFisher Scientific)

Product Name:

TURBO DNA-free Kit

Catalog Number:

AM1907

To rid our RNA preparations of any gDNA contamination and prepare pure samples for cDNA synthesis we utilize Ambion's TURBO DNA-free kit. We selected this kit because of its simple, two step method that never fails to remove the DNA contaminants that carry over into our RNA preparations. We use the DNA-free kit following isolation of RNA using standard phenol-chloroform methods from a whole blood pellet in TRIzol.

Experimental Design and Results Summary

Application

DNA removal from RNA preparations

Starting Material

RNA preparation from blood, tissue

Protocol Overview

Follow the two step protocol as outlined by the manufacturer: 1. Incubate each RNA sample with the DNase enzyme and corresponding buffer. 2. Add the DNase inactivation reagent and incubate, mixing the sample throughout the incubation to ensure even distribution of the reagent throughout each tube. Centrifuge each tube to pellet the reagent so that the DNA-free RNA sample can be aspirated off and transferred to a fresh tube.

Tips

Prepare a master mix when dealing with a large number of samples.

Results Summary

Successful removal of DNA from RNA preparations can be confirmed using a number of methods, including assessment of stable base fluorescence following 30 cycles of qPCR (primers must be designed for your target(s) of interest).

DOI or PMID #

N/A

Additional Notes

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Summary

The Good

Relatively inexpensive, time efficient, simple protocol, reliable.

The Bad

No problems to report.

The Bottom Line

I highly recommend this product as a necessary, important intermediate step between RNA isolation from whole blood and cDNA synthesis. The kit is relatively inexpensive, and with two simple steps it's incredibly easy to use without adding too much time onto the entire reverse transcription process.

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