PCR Product Profiles

Wizard^® SV 96 PCR 
Clean-Up System

Wizard^® SV 96 PCR Clean-Up System is a high-throughput format that integrates the utility of the single, spin column format into a 96-well plate that can be automated on a liquid handling workstation equipped with vacuum manifold and gripper. The system provides a fast, simple technique for efficient recovery of purified DNA fragments generated by PCR amplification. The DNA can be used for automated fluorescent DNA sequencing, cloning, labeling, restriction enzyme digestion, or DNA microarray analysis without further manipulation. The system provides the ability to automate the purification process on any 96-well liquid handler with a gripper. Using this method, double-stranded DNA fragments are purified from 96 samples in approximately 20 minutes. Purification is achieved without phenol/chloroform extraction or ethanol precipitation.

Figure 1. Wizard® SV 96 PCR Clean-Up System protocol overview.

The Wizard^® SV PCR Clean-Up System efficiently purifies PCR fragments from sample reactions using 96-well vacuum filtration steps, eliminating the need for laborious centrifugation. An equal volume of Membrane Binding Solution is added to PCR samples and then applied to the SV 96 Binding Plate. Double-stranded PCR products bind to the SV 96 Binding Plate. Washing of the bound PCR fragments does not require disassembly of the manifold, and filtrate waste products are delivered directly to a vacuum trap, eliminating the need to empty waste collection vessels during purification. Purified PCR products are eluted from the Binding Plate with Nuclease-Free Water and collected into a 96-well Collection Plate for easy storage and direct processing for downstream applications.

FEATURES

Fast: Purify up to 96 PCR reactions in as little as 20 minutes. A simple 3-step protocol is easily implemented on any vacuum equipped automated liquid handler with a gripper.

High Yields: Reproducibly purify a wide range of DNA fragment sizes. Fragments ranging from 100bp-10kb can be isolated with up to 90% recovery.

Figure 2. Percent recovery of purified PCR products. Variable sizes of PCR fragments directly purified from PCR samples. Purified and unpurified PCR products run on 1% agragose gel and stained with EtBr. Percent recovery quantitated by EtBr staining using a Hitachi FMBIO fluorescent scanner (n=6).

Reliable Performance: DNA fragments are purified from a wide variety of amplification conditions including a variety of commonly used DNA Polymerase and PCR co-additives.

Wide Range of DNA Fragment Sizes.

Figure 3. Agarose gel analysis. 2% agarose gel stained with ethidium bromide shows purified (P) and unpurified (U) PCR fragments over a wide range of PCR product sizes.

In the Presence of PCR Amplification Co-additives.

PCR Amplification with a Variety of Polymerase Mixtures.

Versatile Utility: Purified PCR fragments are suitable for a variety of molecular biology techniques including automated fluorescent sequencing and microarray analysis.

Choice of Format: Purify PCR products manually or automate for walk-away purification. Reagent volumes provided are sufficient for automated purification protocols.

Additional Information

DNA and RNA Purification

Wizard is a trademark of Promega Corporation and is registered with the U.S. Patent and Trademark Office.

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Wizard^® SV 96 PCR Clean-Up System