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REPLI-g UltraFast Mini Kit

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).

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

1. Hosono, S. et al. (2003) Unbiased whole-genome amplification directly from clinical samples. Genome Res. 13, 954.

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