QIAGEN Column DNA Extraction From Whole Blood

January 23, 2017

Albert Einstein College of Medicine
Clinical Research
Pre-doctoral researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

QIAGEN

Product Name:

DNeasy Blood & Tissue Kit

Catalog Number:

69504

For our clinical studies, we utilize QIAGEN's DNeasy blood and tissue kit to extract DNA from dried blood spots on Whatman FTA cards/filter papers. The protocol is performed according to the manufacturer's guidelines with minor modifications. The quality and concentration of the DNA extracted from our samples using this kit allow us to carry out many different, downstream experiments, including qPCR assays and genomic sequencing.

Experimental Design and Results Summary

Application

DNA extraction

Starting Material

Human blood (dried spots or liquid), other biological fluids and tissues.

Protocol Overview

Incubate your sample (dried blood spot pieces, fluid) in a series of buffers and enzymes included in the kit (ATL, proteinase K, AL buffer, ethanol -- not included). The temperatures and conditions of each incubation are outlined in the product manual included in the kit. Following these incubations, each sample is washed twice (AW1 and AW2 buffers) with a spin following each wash. The elution (AE) buffer is applied following the wash steps. See 'tips' for suggestions for high yield results.

Tips

Re-use columns to avoid prematurely running out before the kit is finished (dump off filtrate and re-use). If starting from dried blood spots, cut up 3-5 small pieces of the blood on the filter paper and incubate in the first buffer (ATL buffer). For high yield, incubate with elution buffer (AE buffer) for longer than recommended by the manufacturer. Apply half of your desired final volume, incubate, and spin. Repeat with second half of your final desired volume to maximize elution from the column membrane.

Results Summary

Following the final step (elution), measure the DNA concentration and purity in each sample using a NanoDrop.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Cost effective, time efficient, high DNA yield, quality results.

The Bad

Collection tubes often run out before the spin colums (see tips), expensive when compared to traditional DNA extraction methods.

The Bottom Line

I would highly recommend this product for DNA extraction from dried whole blood spots (as well as from biological fluids). The kit is easy to use and yields high concentrations of quality DNA product in a short time frame.

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