Purification of PCR Products

University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Qiagen

Product Name:

QIAquick PCR Purification Kit

Catalog Number:

28104

I have amplified a bacterial gene using PCR which was intended for cloning. I used QIA quick PCR purification kit to get rid of left over primers, dNTPS, and enzyme which otherwise interfere with cloning reaction. My products were of 1Kb length and got good yield after running through this kit. Recovery was spot on and got 85% recovery.

Experimental Design and Results Summary

Application

Purification of PCR products used in cloning reaction.

Starting Material

Bacterial PCR products (specifically lacZ gene product)

Protocol Overview

Part of lacZ gene (E.coli genomic DNA) was PCR amplified (which is of 1kb size) and then purified using QIA quick PCR purification kit. Loaded 50 microliter PCR product into the columns provided in the kit. Then followed easy-to-follow kit instructions. Finally eluted in 30 microliters of molecular biology grade nuclease free water.

Tips

I recommend final elution with 30 microliters of nuclease free water instead of 50 microliters of elution buffer. This will be helpful if your using this product in electro transformation to prevent arching (due to slats in buffer).

Results Summary

Protocol was easy-to-follow. Recovery was 85% of the total input.

Additional Notes

None

Image Gallery

Summary

The Good

Easy-to-follow protocol. Entire protocol will be finished in less than a hour.

The Bad

None

The Bottom Line

Highly recommend this kit if you are a routine cloning molecular biologist.

Join the discussion