Looking Beyond PCR: Isothermal Amplification

Limited sample material resulting in insufficient DNA input is a common hurdle for downstream analysis. This problem can be solved using a variety of methods, including techniques that utilize isothermal amplification. Several methods of isothermal amplification are available – each with advantages and challenges. In this webinar, we will cover two methods, WGA and LAMP, that are commonly used in today’s laboratories.

 

Whole genome amplification (WGA) is a valuable tool to produce large quantities of genomic DNA from limited samples or even single cells. Using a WGA technique called multiple strand displacement amplification (MDA), we will show how the challenges of low DNA input can be overcome using a next generation enzyme, EquiPhi29 DNA Polymerase, an engineered phi29 DNA Polymerase mutant. We will demonstrate that uniform amplification can be obtained from single cells, while leveraging time-saving protocols.

 

Another method of isothermal amplification, loop-mediated isothermal amplification (LAMP), is a rapid method for detecting low amounts of DNA. This technique eliminates the need for a thermal cycler and is increasingly used in rugged field settings for the rapid diagnosis of plant pathogens or infectious disease agents like malaria, Zika, or tuberculosis. We will show how LAMP may be used in specific molecular diagnostics applications.

 

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