Wizard® MagneSil™ PCR Clean-Up System
The Wizard® MagneSil™ PCR Clean-Up System is designed to remove impurities from PCR reactions,
resulting in high quality and yield of double-stranded PCR products. This novel system
removes primers and non-targeted amplification products, such as primer-dimers and excess
nucleotides, from PCR reactions. The
system is
designed to be automated on laboratory robotic systems for unattended
purification.
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Automated purification of PCR
samples on a Beckman Coulter Biomek®
FX with the Wizard® MagneSil™
PCR Clean-Up System. |
PCR
clean-up is performed using Promega’s unique MagneSil™ Paramagnetic Particles
and the MagnaBot® Magnetic Separation Device (Cat.# V8151). This innovative device is designed for use on
robotic platforms.
The MagneSil™ PCR Clean-Up procedure is fast and reliable.
PCR products bind to MagneSil™ Particles in the presence of guanidine
hydrochloride and remain tightly bound during washing.
Purified DNA is eluted in water and may be used for automated fluorescent
sequencing and microarray spotting.
FEATURES
Utilizes MagneSil™ Magnetic Particle Technology
DNA capture
is performed using MagneSil™ paramagnetic
particles. No centrifugation or vacuum required.
Full Automation
The system offers the
distinct advantage of complete walk-away automation. Process multiple plates without
user intervention.
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Automated purification
of PCR products using the MagneSil™ PCR Clean-Up System on the Beckman Coulter Biomek®
FX. |
Qualified for Fluorescent
Sequencing and Microarray Analysis
The
MagneSil™ PCR Clean-Up System purifies PCR products for use in a number of molecular
biology applications including fluorescent sequencing and microarray analysis.
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The
Wizard® MagneSil™ PCR Clean-Up System reproducibly purifies a
1,000bp PCR
product for fluorescent sequencing. The purified PCR product was sequenced using an
ABI PRISM® BigDye™ Terminator Cycle Sequencing Kit. The sequencing reactions were then cleaned up
using the Wizard® MagneSil™ Sequencing Reaction Clean-Up System (Cat.# A1830). The purified PCR product was sequenced using an
ABI PRISM® BigDye™ Terminator Cycle Sequencing Kit. The sequencing reactions were then cleaned up
using the Wizard® MagneSil™ Sequencing Reaction Clean-Up System (Cat.# A1830). |
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Successful microarray production from PCR probes
purified with the Wizard® MagneSil™ PCR Clean-Up System.300bp Kanamycin RT-PCR
products were purified using Wizard®
MagneSil™ PCR Clean-Up System, serially diluted and
spotted in replicate for production of control microarrays. Image represents sample
quadrants of Cy®3 (green) and Cy®5 (red) target hybridizations. The system
provides consistently high yields, spot morphology, and hybridization efficiency. |
Consistent Performance
Remove lower
molecular weight DNA, including primers and primer-dimers, up to 100bp. Primer removal is
consistently greater than 98%.
Recovery of PCR products by size. PCR samples (100µl)
using either PCR Master Mix (Cat.# M7505) or AmpliTaq® DNA Polymerase
(Perkin
Elmer) were purified using the Wizard® MagneSil™ PCR Clean-Up System
manually. The yield of each PCR product was
determined by agarose gel electrophoresis on a 1% agarose gel stained with ethidium
bromide. The gel was imaged and bands were
quantitated using a Molecular Dynamics FluorImager®. These samples consisted of a single size product
with no primer-dimers or primers visible on the gel. The purified PCR products were normalized to an equivalent amount of
starting material. The mean and standard
deviation of 12 individual determinations for each product size is
shown.
Reproducible Results
The Wizard® MagneSil™ PCR Clean-Up System has
been used to purify PCR products from 150bp up to 23kb in size. Demanding molecular biology
applications, such as fluorescent sequencing and microarray analysis, require robust
results from a wide size range of PCR products. Greater than 80% recovery can be obtained with fragments larger than 500bp.
The Wizard®
MagneSil™ PCR Clean-Up System removes primer-dimers from PCR products with high yield
and consistent results. A titration of unpurified prothrombin PCR product
is shown before purification (100%, 50%, 25%, 12.5% and 6.25% of the PCR sample).
The ~100bp primer-dimers are visible in these lanes. Eight replicates purified using
the Wizard® MagneSil™ PCR Clean-Up System show the 506bp prothrombin PCR product with no
visible primer-dimers. The ethidium bromide-stained 1% agarose gel was imaged using
an Alpha Innotech FluorChem™ fluorescence imaging system. Lane M, 100bp DNA
Ladder (Cat.# G2101).
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