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For the fastest reliable whole genome amplification
The REPLI-g UltraFast Mini Kit provides highly uniform whole genome amplification from a variety of sample types in just
6090 minutes. The new REPLI-g UltraFast DNA Polymerase allows fast, accurate amplification of genomes, even from
limited samples, and generates DNA suitable for direct use in a range of genetic analyses.
Benefits of the REPLI-g UltraFast Mini Kit include:
Time savings fast and easy sample amplification in just 6090 minutes
Reliable amplification no change in locus or sequence representation and no amplification in the absence of template
Versatility optimized protocols for different sample types
High-fidelity whole genome amplification (WGA)
REPLI-g UltraFast technology provides fast and highly uniform DNA amplification
across the entire genome, with negligible sequence bias (1). The method
is based on Multiple Displacement Amplification (MDA) technology isothermal
genome amplification utilizing a new, uniquely processive DNA polymerase
that gives DNA products of up to 100 kb in length in a much shorter time
while still avoiding the high sequence bias of PCR-based amplification
methods. In addition, a unique alkaline denaturation buffer guarantees
gentle denaturation of genomic DNA and avoids fragmentation of template
DNA caused by heat denaturation methods. This ensures more sensitive and
more reliable results in downstream assays.
Fast amplification of small samples
Amplification of genomic DNA using the REPLI-g UltraFast Mini Kit involves
3 basic steps (see flowchart). First, the sample (10 ng purified genomic
DNA, 0.5 l whole blood, or 300 tissue culture cells) undergoes
gentle alkaline denaturation, avoiding fragmentation and damage of template
DNA. Next the sample is neutralized, and finally incubated with REPLI-g
UltraFast DNA Polymerase at 30C. The amplified DNA is ready to use after
60 minutes without further purification. The average DNA yield is 10 g
per 20 l reaction after 90 minutes. If time is very limited, sufficient
product is available for downstream genetic analysis after just 45 minutes
(Figure 1).
Reliable control reactions
Negative control samples (without template DNA) are not amplified, providing
a reliable indicator of the presence/absence of sample or contaminating
DNA (Figure 1).
Applications
REPLI-g amplified genomic can be used in a variety of downstream applications,
including:
SNP and STR genotyping
RFLP and Southern blot analysis
Comparative genome hybridization (CGH)
PCR and real-time PCR
Microarray analysis
Reference
1. Hosono, S. et al. (2003) Unbiased whole-genome amplification directly
from clinical samples. Genome Res. 13, 954.
Visit www.qiagen.com/goto/REPLI-g
to discover the benefits of REPLI-g technology today!
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