Kit for isolation of DNA, total RNA and protein

August 21, 2014

Florida State University
Biological Science
Graduate Student
Ethidium bromide stained agarose gel showing total DNA and RNA isolated using kit

Overall

Quality of Results

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Company:

GE Healthcare

Product Name:

Illustra TriplePrep Kit

Catalog Number:

28-9425-44

This kit was used to collect DNA, RNA, and protein for PCR, qPCR, and Western blotting for a circadian rhythms experiment. We thought that having RNA and protein from the same sample would make the results stronger and require less tissue than separately collecting tissue for RNA extraction and additional tissue for protein extraction.

Experimental Design and Results Summary

Application

RNA extraction, protein extraction

Starting Material

Brain tissue, frozen

Protocol Overview

Brain tissue was dissected from anesthetized animals and frozen immediately in a dry ice/ethanol slurry. Tissue was stored at -80 degrees C until extraction. Extraction was performed as per manufacturer instructions.

Tips

None

Results Summary

The RNA extracted with this kit had poor integrity and low concentration. The concentration of RNA was lower than that of competitor kits or TRIzol, and the 260/280 ratio was not good enough for further use. Protein yield was highly variable and less than that collected using a glass homogenizer.

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Summary

The Good

It did get DNA, RNA, and protein out of the same sample.

The Bad

The purity and concentrations were too low to be useful.

The Bottom Line

This kit may perform quite admirably for 2-DIGE and other applications, but it did not perform well for us in extracting high-quality DNA, RNA, or protein, and what it did collect was minute.

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