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Nanoparticle-Assisted PCR (nanoPCR) (CAT#: STEM-MB-0214-WXH)

Introduction

Polymerase Chain Reaction (PCR) is one of the most common technologies used in many laboratories to produce millions of copies of targeted nucleic acid under in vitro conditions. However, PCR faces multiple challenges including limited availability of DNA in the sample, high GC contents of the template, low efficiency, and specificity in amplification. Moreover, some DNA fragments are very difficult to amplify due to their secondary structure and high melting temperature requirement. Nanometer-sized materials (nanomaterials) have offered a possible solution to these problems as these materials have exceptional physio-chemical properties as compared to macroscopic materials. Among these nanomaterials, silicon-based materials, carbon-based materials, semiconductor quantum dots (QDs), and some metals are well-known PCR enhancer. Hence, new PCR has been designed to utilize the unique properties of nanomaterial and is known as nanomaterial-assisted PCR or simply nanoPCR. <br />Nanoparticle associated PCR has advantages of high sensitivity, high specificity and high selectivity, and has been widely used in virus detection and gene sequencing.




Principle

One of the theories involving the gold nanoparticles states that these particles adsorb some of the polymerase and manages the amount of polymerase remaining in the system, which might be necessary in enhancing the specificity of the reaction.
Another theory explains that they adsorb primer pairs and lower the melting temperature at duplex formation between perfectly paired and mispaired primers, which leads to an increase in the specificity of the reaction.

Applications

• Virus detection and gene sequencing
• Diagnosis of diseases
• Detection of defferent pathogens
• The point-of-care (POC) diagnosis of coronavirus disease-19 (COVID-19)
• Long target amplification
• Detection of the telomerase activity
• Genotyping and haplotyping

Procedure

General procedure:
1. Denaturation
2. Annealing
3. Extension

Materials

Carbon nanotubes(CNTs) and Nanopowder(CNP),
Graphene based nanomaterials,
Gold nanoparticles,
Quantum dots,
Magnetic and silica nanoparticles,
Titanium oxide, zinc oxide, and siliver nanoparticles