QIAGEN Kit For DNA Extraction From Agarose Gels

March 01, 2017

Albert Einstein College of Medicine
Microbiology & Immunology
Post-doctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

QIAGEN

Product Name:

QIAquick Gel Extraction Kit

Catalog Number:

28704

QIAGEN's gel extraction kit is useful for extracting and purifying DNA following gel confirmation (band size) of our PCR products of interest. We have used this kit to extract many PCR products of various sizes (up to 10 kb), and the simple procedure consistently provides us with intact, pure products for downstream analysis.

Experimental Design and Results Summary

Application

Cloning and/or sequencing

Starting Material

DNA (up to 10 kb)

Protocol Overview

Use a clean scalpel/knife to cut each PCR product of interest from the gel and subsequently place each gel in a fresh tube. Add 3x volume Buffer QG to each tube. Incubate for 10 minutes at 50C. Once the gel has dissolved, ensure that the color of each sample is yellow. Add 1x volume of isopropanol to each tube and mix. Apply each sample to a column and spin for 1 minute. Discard the flow through and complete the wash steps (i.e. Buffer QG, Buffer PE). Elute the DNA by adding Buffer EB, incubating at room temperature (we incubate for minimum of 5-10 minutes), and centrifuging to collect.

Tips

Volumes added to each tube are dependent on size of gel slice -- try to cut all slices the same size for consistency and simplicity. Use colorless tubes -- must interpret pH based on color.

Results Summary

PCR product quality and concentration can be determined by NanoDrop and confirmed following sequencing analysis.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Easy to use/simple protocol, time effcient, reliable.

The Bad

Like all kits, expensive.

The Bottom Line

QIAGEN's gel extraction kit is the choice kit for extraction and purification of all of our PCR products. We do a lot of cloning and sequencing, and this kit has been consistently

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