PCR Product Profiles

Wizard^® SV Gel and PCR Clean-Up System

The Wizard^® SV Gel and PCR Clean-Up System is designed to extract and purify DNA fragments directly from PCR reactions or from agarose gels. Fragments of 100bp to 10kb can be recovered from standard or low-melt agarose gels in either TAE or TBE buffers. Up to 95% recovery is achieved, depending upon the DNA fragment size. This membrane-based system, which can bind up to 40µg of DNA, allows recovery of isolated DNA fragments or PCR products in as little as 15 minutes. Samples can be eluted in as little as 15µl of Nuclease-Free Water. The purified DNA can be used for automated fluorescent sequencing, cloning, labeling, restriction enzyme digestion or in vitro transcription/translation without further manipulation.

The Wizard^® SV Gel and PCR Clean-Up System efficiently purifies DNA from agarose gels or directly from PCR reactions. An equal volume of Membrane Binding Solution is added to the sample, and then applied to the SV Minicolumn, using either a spin or vacuum format. DNA binds to the SV Minicolumn and impurities are removed using the Membrane Wash Solution. The DNA is then eluted in Nuclease-free Water and can be directly used for a number of downstream applications.

Figure 1. Flow chart of DNA fragment gel purification or direct PCR product purification using the Wizard® SV Gel and PCR Clean-Up System.

The Wizard^® SV Gel and PCR Clean-Up System efficiently purifies DNA from agarose gels or directly from PCR reactions. An equal volume of Membrane Binding Solution is added to the sample, and then applied to the SV Minicolumn, using either a spin or vacuum format. DNA binds to the SV Minicolumn and impurities are removed using the Membrane Wash Solution. The DNA is then eluted in Nuclease-free Water and can be directly used for a number of downstream applications.

FEATURES

Convenient: A single system allows purification directly from PCR reactions or agarose gel slices.

Fast: Purify DNA fragments or PCR products in as little as 15 minutes.

Choice of Format: Choice of spin or vacuum purification formats. No phenol extraction or ethanol precipitation required.

Guaranteed Performance: DNA purified using the Wizard® SV Gel and PCR Clean-Up System is suitable for fluorescent sequencing. Routinely 700 bases with >98% accuracy can be clearly sequenced using DNA purified by this system.

Figure 2. Sequence of a 1,000bp PCR product purified from a 350mg 1% agarose/TAE gel slice using Wizard® SV Gel and PCR Clean-Up System. Sequencing was performed using ABI BigDye® chemistry.

Consistent Recovery

DNA Recovery from Agarose Gels
Using the Wizard^® SV Gel and PCR Clean-Up System, DNA fragments ranging from 100bp to 10kb are purified with excellent recovery rates. Recovery can be achieved with fragments as large as 23kb and with double-stranded DNA as small as 55bp. The SV Gel and PCR Clean-Up System works with standard or low melting point agarose and is compatible with both TAE and TBE buffer systems. The high binding capacity of the SV membrane allows processing of multiple gel slices using a single column.

Figure 3. Gel analysis of 100bp, 500bp, 1,000bp, and 3,199bp DNA fragments before (U; unpurified) and after (P; purified) gel extraction using the Wizard® SV Gel and PCR Clean-Up System. The marker (M) used is the 1kb DNA Ladder (Cat.# G5711).

Table 1. Percent Recovery of DNA Fragments (100bp-9kb) from Agarose Gel Slices Using the Wizard^® SV Gel and PCR Clean-Up System.

DNA Recovery Directly from PCR Reactions
The Wizard^® SV Gel and PCR Clean-Up System effectively purifies a wide range of DNA fragment sizes from PCR. The system effectively removes unincorporated primers from the amplification reaction. Mineral oil does not interfere with purification. In addition, Wizard^® SV chemistry has been shown to be compatible with a variety of PCR additives.

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Wizard^® SV Gel and PCR Clean-Up System