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High-Throughput System Generates Ultra-Pure PCR Products Suitable for All,,,Applications


Simultaneous purification of 96 PCR products

Scott Basehore Jeff Braman
Stratagene

Andrew Firmin
Phenogenex

The StrataPrep 96 PCR purification kit is a new member of the StrataPrep product line. This kit offers a high-throughput format for purifying ultra-pure PCR product DNA from large numbers of samples. The StrataPrep 96 PCR purification kit uses the same silica matrix-binding technology as the other StrataPrep Kit products, but in a 96-well plate format. A vacuum manifold and centrifuge are used to recover 5 g per well of DNA ranging in size from approximately 100 bp to 23,000 bp. Purified PCR products are free of PCR artifacts as well as contaminants from adjacent wells and oil used in some PCR protocols. The purified PCR products are at an ideal concentration for automated fluorescent dideoxynucleotide sequencing. Better sequence data was obtained with a template purified with the StrataPrep 96 PCR kit, compared to the same PCR template with another commercially available kit.

In 1997, Stratagene introduced the StrataPrep PCR purification kit1, an effective miniprep-scale tool for purifying PCR products in applications such as cloning and automated fluorescent dideoxynucleotide sequencing2. The basis of this successful product is its unique glass fiber matrix, which captures DNA in the presence of a chaotropic salt. With the kits wash solution, unincorporated amplification primers and nucleotides, buffer components, enzymes, and nonspecific amplification products smaller than 100 bp can be removed from the matrix-bound PCR products. To recover the desired PCR products, low-ionic-strength buffer is added to the fiber matrix. This step is followed by centrifugation to collect the DNA at a concentration that is suitable for many applications.

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