Biohelix/IsoAmp®III Universal tHDA Kit/H0120S/50 reactions (50 µl reaction)

Description:

   

IsoAmpIII Universal tHDA kit can be used to amplify and detect short DNA sequences (70 bp - 120 bp) at a constant temperature. The kit can be used with a variety of templates, including microbial genomic DNA, viral DNA, plasmid DNA, and cDNA. A single copy of target DNA can be amplified by tHDA and detected by agarose gel electrophoresis when optimized primers and buffer are used.

IsoAmp® III Universal tHDA kit is based on a third-generation tHDA formula for more robust and specific amplification with reduced non-specific products due to primer-dimer. In some cases, rapid amplification may be achieved by adding more IsoAmp® III Enzyme Mix into the reaction (unpublished observation). The volume of IsoAmp® III Enzyme Mix has been reduced to 2 µL per reaction, which allows the user to add more enzymes (2 X or 3 X) for possible high-speed amplification. IsoAmp® III Enzyme Mix (Cat# H1020S) can be purchased separately.

IsoAmp® III Universal tHDA kit supports all reactions which are supported by IsoAmp® II Universal tHDA kit. Selected examples are included in this manual, including end-point tHDA and real-time quantitative HDA (qHDA) in both one-step and two-step formats. A Control Template and a set of specific primers are supplied for the positive control reaction.

 

   

Figure 1. HDA Technology.

   

 

Figure 2.Positive control assays with varying amount of Control Template.
Positive control assays were conducted following the one-step qHDA protocol. Control Template was used at 5 ng - 500 fg per assay in triplicate.

* Negative control reactions may produce primer-dimer, which can be distingushed from specific amplicons by gel-analysis or by melting-curve analysis.

 

Applications:

   

Used to amplify and detect short DNA sequences (80 - 120 bp)

- download Manual

Advantages:
Use of helicase to separate DNA eliminates need for thermocycler
Reactions can be performed at constant temperature.
Works with a variety of templates, including bacterial genomic DNA, viral DNA, plasmid DNA and cDNA.
As low as a single copy of target DNA can be amplified under optimized conditions.
Can be used for real-time quantitative or/and reverse transcription HDA.

     

Kit Components:

   

IsoAmp® III Enzyme Mix (50 reactions)
IsoAmp® dNTP Solution (50 reactions)
10 X Annealing Buffer II
MgSO₄ (100 mM)
NaCl (500 mM)
pCNG1 (Control Template: 1 ng/µl)
NGF3 (Forward Primer: 5 µM)
NGR3 (Reverse Primer: 5 µM)
         

Storage Conditions:

   

Storage Temperature: -20ºC

Recommended storage conditions for the tHDA kit are -20°C for short-term (less than 6 months) storage and -80°C for storage greater than 6 months.          

Quality Control:

 

Quality Assurance Statement:
Kit components were individually tested for optimal performance in tHDA.

         

Legal:

 
Licenses/Patents/Disclaimers:
NOTICE TO PURCHASER: This product is covered by U.S. patents 7,282,328 and 7,662,594 and pending foreign patents. The purchase price of this product includes a limited, non-transferable license for research use only under the patents owned by BioHelix. 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. Further information regarding this product may be obtained by contacting BioHelix at information@biohelix.com.