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Description:

Whole Genome Amplification (WGA) techniques are a useful tool for cancer and genetic research, and typically used to amplify genomic DNA for archiving and down-stream analysis including genotyping, forensics, comparative genomic hybridization, and single cell analysis.

The Rapisome™ pWGA kit non-specifically amplifies total DNA. Unlike other WGA products, the Rapisome™ pWGA kit is based upon the in vitro reconstitution of naturally existing cellular DNA replication system, which performs fast isothermal DNA amplification without the need for thermocycling, prior heat-denaturation, or added primers.

The pWGA technology uses a primase to synthesize primers on-site, generating multiple initiation sites for random, whole genome amplification. Therefore, there is no need to add random primers to the reaction.

The pWGA technology also utilizes a helicase to denature double-stranded template DNA, eliminating the initial heating step that is required by other WGA technologies. Thus, the Rapisome™ pWGA reaction is truly isothermal.

pWGA is also faster than any other commercially available WGA techniques : the reaction time for pWGA is less than one hour. This makes Rapisome™ pWGA attractive technology for use in diagnostic applications more so than other conventional methods.

Applications:

Used to non-specifically amplify total DNA (Fig. 2).
use 10 ng of high-quality DNA template.
approximately 3 - 5 µg yield after 1 hr incubation: Extending the amplification time (2 hrs) may help further increase yields.

Advantages:
No primers: primase synthesizes primers on-site.
No denaturation step: helicase denatures template DNA.
Low bias : uses a natural replisome..
Simple: just add template DNA.
Rapid: 1hr reaction time

Figure 2. Yield of pWGA during 1hr incubation. The Rapisome™ pWGA reactions containing 10 ng human genomic DNA were terminated after a specified incubation time (0, 10, 20 or 60 min), and 5 µl of each 25 µl reaction was analyzed on 1 % agarose gel. The reactions are in triplicate, and lane M shows 500 ng of 1kb DNA ladder (NEB).

Kit Components:

Storage Conditions:

Quality Control:

Quality Assurance Statement:
Kit components were individually tested and confirmed free of endo- and exo-nuclease activity.

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