PCR & qPCR Thermal Cycler

Discover our wide product variety of high-performance PCR as well as highly efficient real-time PCR thermal cyclers for DNA amplification and quantification.

Our PCR and real-time PCR thermal cyclers are also available as fully automated systems for robotic setups.

Throughout the entire lifetime of your instruments, we support you with our application consulting, temperature calibration, maintenance and qualification services.

PCR and qPCR – Effective nucleic acid detection

It's hard to overstate the importance of these "key molecules" to life. After all, these bio-molecules store genetic information and ensure that genetic information is used and passed on within cells. In short: Nucleic acids store the genetic blueprint of all life forms. They allow this blueprint to be turned into proteins for cellular processes.

Knowledge of nucleic acids has also become central to biomedical analytics and molecular biology. In forensic science, medical diagnostics, genetic research and other areas, analysis of nucleic acids allows for many new insights and opens the door to new diagnostic capabilities. The most common methods are PCR and qPCR:

• PCR (polymerase chain reaction) is a procedure for amplifying specific DNA segments by a factor of many millions within a short time. Here, the genetic material passes through repeated cycles of a prespecified temperature program inside a thermal cycler, where DNA polymerase amplifies it exponentially in a chain reaction. PCR technology makes it possible to amplify even the tiniest amounts of DNA. This capability underlies the detection of hereditary diseases, identification of viral infections, verification of genetic fingerprints, and family tree research.
• qPCR (quantitative PCR), or real-time PCR, expands upon the classic PCR method by measuring the DNA level during the amplification process in real-time. Here, fluorescent markers are used to measure the amount of amplified DNA. Thus it is possible to analyze gene expression and track viral load in a sample using qPCR.

PCR and qPCR have become tried and tested methods for high-sensitivity analyses, in particular in applications with very small sample quantities. Unlike sequencing methods, PCR and qPCR often deliver faster results, because they do not require a full sequencing; instead, they target specific DNA segments for amplification and quantification. With cutting-edge, high-throughput machines, laboratories can save valuable time and resources by employing these methods.

Basics of nucleic acid amplification and quantification

To qualitatively and quantitatively analyze miniscule amounts of DNA or RNA, they first have to be amplified. This means multiplying minute traces of nucleic acids so they can be measured in instruments. One of the most well-known nucleic acid amplification methods is the PCR method.

The PCR process involved multiple steps:

1. Denaturation: The DNA is heated to a high temperature, causing the two strands to separate, similar to a zipper opening. This grants access to the individual strands for what comes next.
2. Primer annealing: After cooling down, short pieces of DNA called primers bind to the ends of the stretch of DNA being amplified. These primers function as starting points for DNA synthesis and set the target region for amplification.
3. Extension: A special molecular machine in the form of an enzyme, DNA polymerase, attaches new DNA building blocks to the primer and thereby begins to replicate the missing strands of DNA. This duplicates the DNA.

In modern laboratories, the PCR process described here runs on thermal cyclers. These are machines that run the temperature-sensitive PCR reactions precisely and without human intervention. The thermal cycler's heating element controls the different temperatures and time intervals in each phase of the PCR process. First, the denaturation process uses a high temperature of 94 °C to 98 °C to separate the DNA strands from each other. For primer annealing, the temperature is then lowered to between about 50 °C to 65 °C before synthesis of new DNA strands takes place at approximately 72 °C.

The chain reaction passes through 20 to 40 cycles, with the amount of DNA doubling each cycle. The DNA fragment can then be detected using various methods such as gel electrophoresis. Since detection only takes place after the amplification process, the method is also known as endpoint PCR.

Cutting-edge PCR thermal cyclers like the Biometra TAdvanced Series in labs today achieve extremely high temperature homogeneity throughout the entire process so that PCR reactions can be reproduced reliably. 

Quantitative PCR (qPCR) is an outgrowth of the PCR method. Unlike endpoint PCR, qPCR can measure the amount of DNA in real-time. For that reason, the method is also known as real-time PCR. It has the advantage that changes in gene expression or viral load in samples can be monitored continuously. In qPCR, special fluorescent dyes (probes) are added and bind to the DNA during amplification of the nucleic acids. The resulting fluorescent signals can be measured each cycle using a detector. The more DNA is produced, the stronger the signal. Thanks to this method, it is possible to detect how many nucleic acids were present in the sample at the beginning. Thus, for example, it is possible to detect the viral load in a coronavirus test.

Labs today use both manual as well as fully automatic real-time PCR thermal cyclers. In addition to high temperature precision, these machines also offer high detection accuracy along with quantification of the nucleic acid concentration. The proven qTOWER Series from Analytik Jena impresses with flexible configuration options – from a choice of consumables to customization of the color module. This versatility makes the qTOWER series the ideal choice for highly diverse quantitative PCR applications.

Applications of PCR and qPCR methods

PCR and qPCR technology is widespread today in many branches of science, research and diagnostics, as well in agricultural and environmental analysis applications. As a highly sensitive and reliable procedure, this method has become established in the following applications, to name only a few:

• Clinical diagnostics: In clinical diagnostics, for example, PCR and qPCR technology supports detection of pathogens like viruses (such as SARS-CoV-2) or bacteria (such as Mycobacterium tuberculosis). The method is also suitable for diagnosing genetic defects like cystic fibrosis and Huntington's disease.
• Cancer research: In cancer research, thermal cyclers play a role in identifying genetic mutations associated with cancer, such as BRCA1/2 mutations.
• Forensics: PCR methods are a standby in crime scene analysis. Thermal cyclers make it possible to identify people through DNA analysis of biological clues like hair, saliva or blood.
• Agriculture: A typical agricultural application for PCR methods might be breeding new plants – here, the method helps identify genes responsible for certain traits. 

Besides the applications listed here, there are other areas where the advantages of the PCR method are brought to bear, for instance when detecting and identifying microorganisms in food or when researching the effect of new medications on gene expression in pharmaceutical research.