Direct-Zol RNA MiniPrep Kit -- A Reliable, Good-Value Kit For RNA Isolation

August 04, 2016

Duke University
Immunology
Graduate student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Zymo Research

Product Name:

Direct-zol™ RNA MiniPrep Kit

Catalog Number:

R2052

Our lab uses this kit to isolate mRNA from plasmablasts cultured in vitro. We disrupt the cultured cells in TRIzol, use this kit to purify RNA, then convert the mRNA to cDNA and amplify the V(D)J rearrangements by PCR, followed by DNA sequencing.We have used hundreds of columns (and corresponding reagents) from this kit, and continue using it because of its value, its ease of use, and the highly reliable performance.

Experimental Design and Results Summary

Application

RNA isolation from cultured cells

Starting Material

Cultured B cells dissociated in TRIzol reagent

Protocol Overview

Cultured cells are disrupted directly in TRIzol reagent. The TRIzol is applied directly to the columns, and the columns are washed several times per the manufacturer's instructions. Total RNA is eluted in water.

Tips

The size of the columns and collection tubes makes it challenging to fully load and unload a standard tabletop microcentrifuge.

Results Summary

We never directly analyze the quality of the purified RNA -- we go directly to cDNA synthesis and downstream PCR. We can amplify V(D)J rearrangements from >90% of our starting samples, but it's not clear whether the loss represents poor quality purified RNA or failed downstream PCR (likely the latter).

Additional Notes

None

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Summary

The Good

Straightforward protocol; reliable performance; good value

The Bad

Difficult to load a microcentrifuge with the maximum number of tubes, due to the slightly oversized columns/collection tubes.

The Bottom Line

Our lab has used >1000 columns and associated reagents from these kits, and we're happy with them. We will keep stocking them for the foreseeable future.

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