HOME ABOUT US TECHNOLOGY PRODUCTS CONTACT US  
 
       
 
 
 
   
  Products
  Diagnostics Products
  Isothermal DNA Amplification
  Primer Navigator™ series
  Helicases
 
 
Rapisome™ pWGA Kit
 

Products - Isothermal DNA Amplification

Catalog #
Size
Concentration
Price
H0300S
25 reactions (25µl reaction)
Discontinued

Download: Manual ; MSDS : Publication

   

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.

  Figure 1. pWGA Technology.


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:

   


pWGA Master Mix: 25 reactions (5 reactions/tube)
Control Template: 20 µl (Human genomic DNA, 5 ng/µl)


   
     

Storage Conditions:

   


Storage Temperature:
-70ºC

     
     

Quality Control:

   


Quality Assurance Statement:

Kit components were individually tested and confirmed free of endo- and exo-nuclease activity.

     
     

Legal:

   

Licenses/Patents/Disclaimers:
NOTICE TO PURCHASER: This product is covered by pending U.S. and foreign patent applications exclusively licensed by BioHelix Corporation from Harvard Medical School. The purchase price of this product includes a limited, non-transferable license for research use only. No other license under these patents is conveyed expressly or by implication to the purchaser by the purchase of the product. The product is not to be used for diagnostic purposes nor is it intended for human use. This product may not be resold, modified for resale, or used to manufacture commercial products without written approval of BioHelix Corp.

 
   
Privacy, Limitation, Warranty, Disclaimer