QuickClean 5M PCR Purification Kit From GenScript Corporation

The QuickClean 5M PCR Purification Kit from GenScript Corporation (Piscataway, NJ) is designed to remove primers, salts, nucleotides, enzymes, and dyes from reactions to allow the DNA of interest to be used in a variety of downstream applications. Although procedures such as PCR and restriction digests are frequently used simply as analytical tools, they also have wide applicability as preparative procedures. In these cases the success of downstream applications, such as cloning, sequencing, or in vitro transcription, can be critically compromised by the presence of unwanted reaction components from preceding procedures. In most cases, the separation of the desired DNA from unwanted contaminants is accomplished by precipitation, affinity resins, or gel purification. The method employed in the QuickClean Kit, a spin column affinity resin, is arguably the most common method used by kit manufacturers.

The QuickClean DNA cleanup protocol is fairly standard for this type of kit and consists of adding a binding solution to the reaction mixture containing the DNA of interest, transferring the mixture to a spin column, subjecting the column to a brief centrifugation, washing away nonbinding components, and eluting the DNA, hopefully free of contaminants. According to the manufacturer, each spin column in the QuickClean Kit can bind up to 20 µg of single- or double-stranded DNA, 70 bp to 40 kb in length. I used the kit to purify a ~650 bp amplicon from a 50 µl PCR reaction, eluted the DNA in 50 µl elution buffer, and ran 10 µl of the eluted DNA next to 10 µl of a duplicate PCR reaction. In the gel image shown, with the sample from the PCR reaction on the left and the cleaned up sample on the right, it is evident that recovery was excellent. I tested the purity of the cleaned-up product by having the amplicon sequenced and was able to read the entire amplicon; 621 bases of very good quality sequence.

The QuickClean 5M PCR Purification Kit is available in a 250 prep package and is quite a bargain compared to the same sized package from one of the leading kit manufacturers.