Rapid Concentration and Purification of Dilute PCR Products

February 11, 2020

University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

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Company:

Promega

Product Name:

ReliaPrep DNA Clean-up and Concentration System

Catalog Number:

A2891

It is very critical to remove unused primers and dNTPs from the final PCR reactions and also concentrate the DNA to high concentration for applications such as high efficiency transformations and restriction cloning reactions. To this end, I used ReliaPrep DNA Clean-up and Concentration kit to concentrate and purify 300 microliter PCR reaction (10.50 microgram DNA) in single Reliaprep purification microcolumn. Finally eluted with 20 microliter nuclease free water with recovery of 80 % and yield of 7.82 microgram.

Experimental Design and Results Summary

Application

Rapid concentration and purification of dilute PCR reactions

Starting Material

300 microliter of pooled PCR reactions representing aroudn 10.5 microgram DNA per Reliaprep microcolumns

Tips

For eluting the DNA inserts grater than 5 kb and upto 10 kb, increase eultion by allowing the columns to stand for 5 minutes before centrifugation to elute.

Results Summary

Total of 10.5 microgram of pooled PCR reactions in a 300 microliter were used for purification and concentration to a volume of 20 microliter in nuclease free water. The standard easy-to-perform protocol was followed according to Kit's instructions. After purification and concentration, the quality and yield was determined by Nanodrop quantification. The yield was 80 % recovery (7.82 microgram in 20 microliter elution) with quality i.e. 260:280 ratio of 1.85 and 260:230 ration of 1.92

DOI or PMID #

N/A

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Summary

The Good

Rapid and fast method. You can purify and concentrate 6 reaction in 10 minute time.

The Bad

None

The Bottom Line

Highly recommended kit for rapid concentration and purification of dilute DNA samples like PCR reactions or Restriction digested DNA samples.

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