Download: Manual ; MSDS

Description:

IsoAmpII 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.

IsoAmpII Universal tHDA kit is based on a second-generation thermophilic Helicase-Dependent Amplification platform. The reactions supported by IsoAmp® II Universal tHDA kit include tHDA, reverse transcription HDA (RT-HDA), real-time quantitative HDA (qHDA) and real-time quantitative RT-HDA (qRT-HDA), from a single reaction buffer.

IsoAmpII Universal tHDA kit contains an enzyme mix, dNTP solution and buffer, which allows flexibility in reaction setup. A Control Template and a set of specific primers are supplied for the positive control reaction.

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)

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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® 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)

Note: Control Template and Primers have been changed.

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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