What is PCR?
Polymerase Chain Reaction (PCR) is a method used to multiply specific DNA fragments exponentially through a series of temperature change cycles. One of the advantages of PCR is the speed and simplicity of the technique. The PCR process can be carried out in a short time, and sample preparation in large quantities can be done easily. Apart from that, PCR also has high accuracy and specificity in detecting DNA so it is very commonly used in various research, especially in the fields of biotechnology and molecular biology.
How does PCR work?
PCR consists of three main process stages. The first stage is Separation or Denaturation, where the template DNA molecule or DNA sample containing the target sequence is heated at a high temperature (usually 94–98°C) so that the DNA strand will open into two strands. The next stage is sticking or annealing. The template DNA strand will bind to the specific primer added due to a decrease in reaction temperature (usually 55–65°C). A primer is a short single DNA fragment or oligonucleotide that contains the nucleotide sequence of a specific target. The next stage is Lengthening or Extension. At this stage the temperature is increased again (usually 75–80°C), so that the DNA Polymerase enzyme will multiply the DNA fragments that have bound to the primer. These stages are repeated several cycles or rounds resulting in complete synthesis and multiplication of new DNA.
PCR application
The use of PCR is very common in various application fields such as molecular clinical diagnostics, molecular biology, food testing and forensics. In molecular clinical diagnostic testing, PCR is used for the detection of specific target genes and identifying genetic diseases, infections and genetic mutations, including infectious diseases such as HIV, HPV, COVID-19, Hepatitis and tuberculosis. In the forensic field, PCR is used to identify DNA profiles. Apart from that, PCR also plays an important role in food safety testing such as pork DNA analysis to ensure the halalness of a product or detection of pathogens related to food safety to ensure products are free from contamination.
Nippon Genetics Europe: FastGene® qFYR Real-Time PCR
To meet the needs of DNA analysis using PCR in your laboratory, we provide FastGene® qFYR Real-Time PCR which comes from Germany and is designed according to laboratory standards so that it will provide accurate test results. This tool can detect quickly using two methods, namely dye and probe based. FastGene® qFYR Real-Time PCR is a multiplex PCR that can detect 4+1 channels for different DNA targets, including additional FAM/SYBR channels for fast dual-mode melt curve analysis. Our real-time PCR has a 96 well plate format which is flexible to use using well plates, PCR tubes, or PCR strip tubes. In addition, the thermal block is designed with holes with a gradient function so that heating and cooling will be faster. FastGene® qFYR Real-Time PCR provides innovative technology with a unique optical detection system combining a high-quality PMT (photomultiplier tube) with a short Fresnel lens to increase signal sensitivity.
FastGene® qFYR Real-Time PCR can analyze the results of various tests such as gene expression, genotyping, HRM (high resolution melt), gene mutation analysis, pathogen detection, GMO detection, protein stability screening, miRNA analysis, and melt curve analysis. PCR results are analyzed using easy-to-use software. In addition, qFYR is also equipped with an Integrated Precision Melt Analysis Tool for probe-based allele discrimination.
Source :
https://www.ncbi.nlm.nih.gov/probe/docs/techpcr/
Wages Jr., J. M. (2005). Polymerase Chain Reaction. In Palmetto Consulting & Research, Tupelo, MS, USA (Ed.), Elsevier Ltd. All Rights Reserved (pp. 243-250).
