PCR Machine- Principle, Parts, Steps, Types, Uses, Examples

PCR machines, also known as thermal cyclers or DNA amplifiers, are laboratory devices that are used to amplify segments of DNA or RNA chosen from the genome with a primer . PCR stands for polymerase chain reaction, a technique that was invented by Kary Mullis in 1983 and revolutionized molecular biology. PCR machines allow scientists to make millions of copies of a specific DNA or RNA sequence in a matter of hours, which enables them to study the genetic material in detail .

PCR machines work by cycling through three main steps: denaturation, annealing, and extension . In denaturation, the DNA template is heated to separate the two strands. In annealing, short synthetic oligonucleotides called primers bind to the complementary regions of the single-stranded DNA. In extension, a heat-stable enzyme called Taq polymerase adds nucleotides to the 3` end of the primers, synthesizing new strands of DNA that are identical to the template . These steps are repeated for 20 to 40 cycles, resulting in an exponential amplification of the target DNA or RNA sequence .

PCR machines require five essential reagents for the reaction: DNA template, Taq polymerase, primers, deoxyribonucleotide triphosphates (dNTPs), and PCR buffer. The DNA template is the source of the target sequence that needs to be amplified. The Taq polymerase is a thermostable enzyme that can withstand high temperatures and catalyze the DNA synthesis. The primers are short sequences of nucleotides that are designed to match the ends of the target sequence and provide a starting point for the Taq polymerase. The dNTPs are the building blocks of DNA that are added by the Taq polymerase to extend the primers. The PCR buffer provides the optimal conditions for the reaction, such as pH, salt concentration, and magnesium ions.

PCR machines consist of three main parts: a thermal block, a heated lid, and a control panel . The thermal block has holes where tubes or plates holding the reaction mixtures can be inserted. The thermal block can change the temperature rapidly and accurately according to a programmed protocol. The heated lid presses against the tops of the tubes or plates and prevents condensation of water from the reaction mixtures on the inside of the lids. The control panel has a graphical display and a key-pad that allow users to enter and monitor the parameters and status of the reaction .

PCR machines have many types and applications in various fields of science and medicine. Some examples of types of PCR are:

• Real-time PCR or quantitative PCR (qPCR), which measures and quantifies the amount of amplified DNA or RNA in real-time using fluorescent dyes or probes .
• Reverse-transcriptase PCR (RT-PCR), which converts RNA to DNA using an enzyme called reverse transcriptase and then amplifies the resulting complementary DNA (cDNA) .
• Nested PCR, which reduces the chances of unwanted products by performing a second PCR using new primers that are nested within the first ones .
• Hot start PCR, which uses antibodies or chemical modifications to inactivate Taq polymerase until a high temperature is reached, preventing nonspecific amplification at low temperatures .
• Multiplex PCR, which amplifies several fragments in a single DNA sample by using multiple sets of primers .
• Long-range PCR, which uses a combination of polymerases and additives to amplify larger fragments of DNA than conventional PCR .
• In situ PCR, which performs PCR directly in cells or fixed tissue on a slide .
• Asymmetric PCR, which amplifies one strand of the target DNA more than the other by using unequal amounts of primers .
• Assembly PCR, which uses overlapping primers to create longer DNA fragments from shorter ones .

Some examples of applications of PCR are:

• Gene expression analysis, which studies how genes are turned on or off in different cell types, tissues, or conditions using RT-PCR or qPCR .
• Genotyping, which detects variations in alleles or genes among different cells or organisms using PCR with specific primers or probes .
• Cloning and mutagenesis, which introduces new DNA sequences into bacteria or other hosts using PCR-generated fragments as inserts for vectors such as plasmids or phages .
• Sequencing, which determines the order of nucleotides in DNA using PCR to prepare and process template DNA for various methods such as Sanger sequencing or next-generation sequencing (NGS).
• Medicine and biomedical research, which uses PCR to diagnose genetic diseases, identify infectious agents, perform prenatal testing, screen embryos for IVF, and more.
• Forensic science, which uses PCR to analyze DNA evidence from crime scenes or paternity tests.
• Environmental microbiology and food safety, which uses PCR to detect pathogens or contaminants in water, soil, food, or other samples.

In summary, PCR machines are powerful and versatile tools that can amplify small amounts of DNA or RNA for various purposes. They work by cycling through three steps: denaturation, annealing, and extension. They require five reagents: template DNA/RNA , Taq polymerase , primers , dNTPs , and buffer . They consist of three parts: thermal block , heated lid , and control panel . They have many types and applications in different fields of science and medicine.

The principle of PCR is based on the natural process of DNA replication, in which a DNA polymerase enzyme synthesizes new strands of DNA complementary to a template strand. However, PCR is carried out in vitro, or in a test tube, using a small amount of template DNA, synthetic DNA primers, free nucleotides, and a thermostable DNA polymerase. The primers are short sequences of nucleotides that are designed to match the ends of the target region of DNA that needs to be amplified. The thermostable DNA polymerase is an enzyme that can withstand high temperatures and catalyze the addition of nucleotides to the growing DNA strands.

PCR consists of three main steps that are repeated for a number of cycles:

• Denaturation: The template DNA is heated to about 94°C to separate the two strands and expose the bases.
• Annealing: The temperature is lowered to about 50-65°C to allow the primers to bind to their complementary sequences on the template strands. This provides a starting point for the DNA polymerase to extend the new strands.
• Elongation: The temperature is raised to about 72°C, which is the optimal temperature for the DNA polymerase to work. The enzyme adds nucleotides to the 3` end of each primer, following the base-pairing rules of A with T and G with C. As a result, two new strands of DNA are formed that are identical to the target region.

The products of one cycle serve as the templates for the next cycle, doubling the amount of DNA with each cycle. After 20-40 cycles, millions or billions of copies of the target region are generated. The amplified DNA can then be used for various purposes, such as sequencing, cloning, diagnosis, or analysis.

A PCR machine, also known as a thermocycler, is a device that performs PCR by cycling the temperature of the reaction mixture according to a programmed protocol. A typical PCR machine consists of the following parts:

• Thermal block: This is the part of the PCR machine that holds the reaction tubes or plates and changes the temperature according to the PCR protocol. The thermal block can have different formats, such as 96-well, 384-well, or 0.2 ml tubes. The thermal block can also have different zones that allow for temperature variation across the block, which is useful for gradient PCR or optimization experiments. The thermal block is usually made of metal, such as silver or aluminum, which have high thermal conductivity and can achieve fast and uniform heating and cooling.

• Heated lid: This is the part of the PCR machine that covers the thermal block and presses against the reaction tube or plate lids. The heated lid prevents condensation of water from the reaction mixtures on the insides of the lids, which could affect the volume and concentration of the PCR components. The heated lid also ensures a tight seal between the tube or plate and the thermal block, which improves thermal contact and reduces evaporation. The heated lid can be adjusted for different heights and pressures of the tubes or plates.

• Control panel: This is the part of the PCR machine that allows the user to program and monitor the PCR protocol. The control panel usually has a graphical display that shows the current status of the PCR machine, such as temperature, time, cycle number, and error messages. The control panel also has a keypad or a touch screen that allows the user to enter and edit the PCR protocol parameters, such as temperature, time, ramp rate, number of cycles, and hold steps. Some PCR machines also have a USB port or a WiFi module that enables data transfer and remote control.

• Air vents: These are the parts of the PCR machine that facilitate air circulation and cooling of the device. The air vents are located on the front, lateral, and bottom sides of the PCR machine and allow air intake and output. The air vents also prevent overheating and damage to the internal components of the PCR machine.

These are the main parts of a PCR machine that enable it to perform PCR efficiently and accurately. By understanding how each part works, you can better troubleshoot any problems that may arise during your PCR experiments.

PCR components are the chemical ingredients that are required for a PCR reaction to take place. They include:

• DNA template: This is the sample DNA that contains the target region to be amplified. The DNA template can be extracted from various sources, such as blood, saliva, hair, tissue, etc. The quality and quantity of the DNA template can affect the efficiency and accuracy of PCR.
• DNA polymerase: This is the enzyme that synthesizes new DNA strands complementary to the template strands. The DNA polymerase used in PCR must be heat-resistant, as it has to withstand repeated cycles of high temperature denaturation. The most commonly used DNA polymerase is Taq polymerase, which is derived from a thermophilic bacterium called Thermus aquaticus. Other DNA polymerases, such as Pfu polymerase and Phusion polymerase, have higher fidelity and can produce longer amplicons than Taq polymerase .
• Primers: These are short pieces of single-stranded DNA (usually 20-30 nucleotides long) that are designed to match the sequences at the ends of the target region. Primers provide the starting point for DNA polymerase to extend new DNA strands. Two primers are used in each PCR reaction: a forward primer that binds to the 3` end of the template strand, and a reverse primer that binds to the 5` end of the complementary strand. Primers should be specific, stable and complementary to the template .
• Nucleotides: These are the building blocks of DNA, consisting of four types: adenine (A), thymine (T), guanine (G) and cytosine (C). Nucleotides are supplied as deoxyribonucleotide triphosphates (dNTPs) in PCR, which provide the energy and the base for DNA polymerase to add to the growing DNA strand .
• Reaction buffer: This is a solution that provides the optimal conditions for PCR, such as pH, salt concentration and cofactors. The reaction buffer usually contains Tris-HCl (to maintain pH), potassium chloride (KCl) (to stabilize DNA polymerase), magnesium chloride (MgCl2) (to serve as a cofactor for DNA polymerase) and sometimes additives such as bovine serum albumin (BSA) (to prevent nonspecific binding) or dimethyl sulfoxide (DMSO) (to enhance primer annealing) .

These components are mixed together in a small tube or well and subjected to repeated cycles of heating and cooling in a PCR machine or thermocycler. Each cycle consists of three steps: denaturation, annealing and extension. Denaturation involves heating the mixture to 94°C to separate the double-stranded DNA template into single strands. Annealing involves cooling the mixture to 50-65°C to allow the primers to bind to their complementary sequences on the template strands. Extension involves raising the temperature to 72-80°C to enable DNA polymerase to synthesize new DNA strands from the primers .

By repeating these cycles, the target region of DNA is exponentially amplified, resulting in millions or billions of copies that can be used for further analysis or applications.

PCR is a cyclic process that consists of three main steps: denaturation, annealing, and elongation. These steps are repeated for a number of cycles, usually 25 to 40, to amplify the target DNA sequence exponentially. Each cycle takes about 1 to 2 minutes to complete, depending on the length of the target sequence and the type of PCR machine used. The following is a brief description of each step:

• Denaturation: This step involves heating the reaction mixture to a high temperature, usually around 94°C, for a short period of time, usually 15 to 30 seconds. The high temperature breaks the hydrogen bonds between the complementary strands of the template DNA, resulting in the separation of the double-stranded DNA into single-stranded DNA molecules. This step prepares the template DNA for the next step of annealing.

• Annealing: This step involves cooling down the reaction mixture to a lower temperature, usually between 50°C and 65°C, for another short period of time, usually 20 to 40 seconds. The lower temperature allows the primers, which are short synthetic oligonucleotides that are complementary to specific regions of the template DNA, to bind or anneal to their respective target sequences on the single-stranded DNA molecules. The primers serve as the starting points for the DNA polymerase enzyme to synthesize new DNA strands in the next step of elongation.

• Elongation: This step involves raising the temperature again to an optimal level for the DNA polymerase enzyme to function, usually around 72°C, for a longer period of time, usually 30 seconds to 2 minutes depending on the length of the target sequence. The DNA polymerase enzyme adds nucleotides to the 3` end of each primer, following the base-pairing rules of A with T and G with C, and extends the primer into a new complementary DNA strand that matches the template DNA strand. This step results in the formation of two new double-stranded DNA molecules that contain the target sequence.

After one cycle of PCR, the amount of target DNA sequence is doubled. After two cycles, it is quadrupled. After three cycles, it is octupled. And so on. By repeating these three steps for multiple cycles, PCR can amplify a very small amount of target DNA sequence into millions or billions of copies in a matter of hours.

The PCR machine operating procedure involves the following steps:

1. Prepare the PCR tubes or plates with the required reagents, such as DNA template, primers, dNTPs, buffer, and DNA polymerase. Label the tubes or plates with the sample ID and the PCR program name.
2. Load the tubes or plates into the PCR machine or thermal cycler. Make sure they are tightly sealed and fit well into the slots. If the machine does not have a heated lid, add mineral oil to prevent evaporation.
3. Set up the PCR program on the machine according to the primer and template parameters. The program should include three main steps: denaturation, annealing, and extension. Each step should have a specific temperature and duration. The program should also specify the number of cycles to repeat these steps.
4. Start the PCR program and monitor the progress on the machine display. The machine will automatically change the temperature and time for each step and cycle.
5. When the PCR program is finished, remove the tubes or plates from the machine and store them at 4°C or -20°C until further analysis.
6. Evaluate the amplified DNA by agarose gel electrophoresis followed by ethidium bromide staining or other methods.

Some examples of PCR machines are:

• Biometra TAdvanced Thermal Cycler Series (Analytik Jena)
• MiniAmp™ Plus Thermal Cycler (ThermoFisher Scientific)
• Esco Swift Thermal Cyclers
• GET-S SERIES THERMAL CYCLER (Bio-gener)
• MiniAmp Plus Thermal Cycler and the MiniAmp Thermal Cycler (Delta Science)

These machines have different features and specifications, such as sample capacity, ramping rate, temperature control, touch-screen interface, WiFi connectivity, email notification, storage capacity, etc.

PCR is a versatile technique that can be modified and adapted for different purposes and applications. Depending on the specific goal and requirement of the experiment, different types of PCR can be used to achieve the desired outcome. Some of the common types of PCR are:

• Real-Time PCR (quantitative PCR or qPCR): This type of PCR allows the detection and quantification of the amplified DNA in real-time by using fluorescent dyes or probes that bind to the DNA and emit signals. The amount of fluorescence is proportional to the amount of DNA present in each cycle, and a threshold cycle (Ct) value can be calculated to determine the initial amount of template DNA in the sample. Real-time PCR is useful for gene expression analysis, genotyping, mutation detection, and pathogen identification.
• Reverse-Transcriptase PCR (RT-PCR): This type of PCR uses an enzyme called reverse transcriptase to convert RNA into complementary DNA (cDNA), which can then be amplified by PCR. RT-PCR is commonly used to measure the expression level of specific genes by using mRNA as the template. RT-PCR can also be used to detect RNA viruses, such as HIV and SARS-CoV-2.
• Multiplex PCR: This type of PCR uses multiple pairs of primers to amplify different target regions simultaneously in a single reaction. Multiplex PCR can save time, cost, and sample material by reducing the number of reactions needed. Multiplex PCR can be used for various applications, such as detection of multiple pathogens, identification of genetic markers, and forensic analysis.
• Nested PCR: This type of PCR involves two rounds of amplification using two sets of primers. The first set of primers amplifies a broad region of interest, and the second set of primers amplifies a smaller region within the first product. Nested PCR can increase the specificity and sensitivity of PCR by reducing nonspecific amplification and background noise. Nested PCR can be used for detection of low-abundance targets, such as rare mutations or pathogens.
• High-Fidelity PCR: This type of PCR uses a DNA polymerase enzyme that has a high accuracy and low error rate during DNA synthesis. High-fidelity PCR can minimize the introduction of mutations or errors in the amplified DNA, which is important for applications that require precise sequence information, such as cloning, sequencing, and gene editing.
• Fast PCR: This type of PCR uses a fast-cycling protocol that reduces the time required for each cycle by optimizing the temperature and duration of denaturation, annealing, and extension steps. Fast PCR can also use a fast DNA polymerase enzyme that has a high processivity and catalytic rate. Fast PCR can shorten the overall run time of PCR and increase the throughput of samples.
• Hot-Start PCR: This type of PCR uses a modified DNA polymerase enzyme that is inactive at room temperature and becomes active only at high temperature. Hot-start PCR can prevent nonspecific amplification and primer-dimer formation that may occur during the initial setup of the reaction. Hot-start PCR can improve the specificity and yield of PCR.

PCR has a broad range of applications, not only in basic research but also in the areas of medical diagnostics, forensics, and agriculture. As described below, some examples of PCR applications include:

• Gene expression: PCR can be used to measure the levels of mRNA for a specific gene in different cell types, tissues, or organisms at a given time point. This can reveal how genes are regulated and how they respond to various stimuli or conditions. To perform gene expression analysis by PCR, RNA is isolated from the samples of interest and reverse-transcribed into cDNA. The amount of cDNA amplified by PCR reflects the original levels of mRNA for the target gene .
• Genotyping: PCR can be used to detect sequence variations in alleles in specific cells or organisms. For instance, PCR can be used to identify transgenic organisms, such as knockout or knock-in mice, by designing primers that flank the regions of interest and assess genetic variations based on the presence or absence of an amplicon and/or its length. PCR can also be used to screen for genetic mutations or polymorphisms associated with diseases or traits in humans or other species.
• Cloning: PCR can be used to amplify a DNA fragment of interest from a template DNA source, such as genomic DNA, cDNA, or plasmid DNA. The amplified DNA fragment can then be inserted into a vector for further manipulation, such as sequencing, expression, or functional analysis. PCR cloning is faster and more convenient than traditional cloning methods that rely on restriction enzymes and ligases.
• Mutagenesis: PCR can be used to introduce specific changes in a DNA sequence, such as point mutations, insertions, or deletions. This can be achieved by using primers that contain the desired mutation and performing an overlap extension PCR. Alternatively, PCR can be combined with site-directed mutagenesis methods that use synthetic oligonucleotides to anneal and replace a target region in a plasmid. Mutagenesis by PCR can be useful for studying the function and regulation of genes or proteins.
• Methylation analysis: PCR can be used to detect and quantify the methylation status of specific DNA regions, such as promoters or CpG islands. Methylation is a type of epigenetic modification that affects gene expression and is involved in various biological processes and diseases. To perform methylation analysis by PCR, DNA is treated with bisulfite, which converts unmethylated cytosines to uracils, while leaving methylated cytosines unchanged. The bisulfite-treated DNA is then amplified by PCR using primers that discriminate between methylated and unmethylated alleles.
• Sequencing: PCR can be used to prepare DNA samples for sequencing analysis, which can reveal the exact nucleotide sequence of a DNA region of interest. Sequencing can be used for various purposes, such as identifying mutations, comparing genomes, or analyzing gene expression. PCR can be used to amplify the target DNA region before sequencing, as well as to add sequencing adapters and barcodes for multiplexing. PCR is also involved in next-generation sequencing (NGS) methods that use high-throughput platforms to sequence millions of DNA fragments simultaneously.

• Medical, forensic, and applied sciences: PCR can be used for various applications that have practical implications for human health and society. For example, PCR can be used to diagnose infectious diseases by detecting the presence of pathogens in biological samples. PCR can also be used to test for genetic diseases or predispositions by analyzing the DNA of patients or their relatives. Furthermore, PCR can be used for forensic purposes by identifying individuals based on their DNA profiles or fingerprints. Additionally, PCR can be used for agricultural purposes by detecting genetically modified organisms (GMOs) or pests in crops.

  PCR Advantages and Limitations

PCR is a powerful and versatile technique that has many applications in biology and medicine. However, it also has some limitations and challenges that need to be considered. Here are some of the advantages and limitations of PCR:

Advantages

• High specificity: PCR can distinguish DNA sequences by just one nucleotide, making it a very accurate technique. It can also use specific primers that target only the region of interest, reducing the chances of amplifying unwanted DNA .
• High sensitivity: PCR is a very useful technique when the amount of DNA sample is limited because it allows the detection of even a single copy of a specific DNA template . It can also amplify DNA from degraded or contaminated samples, such as forensic or archaeological specimens.
• High speed: PCR can produce millions or billions of copies of a DNA region in a matter of hours, making it much faster than conventional cloning methods . It can also be automated and performed in parallel, increasing the throughput and efficiency of the analysis.
• High versatility: PCR can be used for a variety of purposes, such as gene expression, genotyping, sequencing, cloning, mutagenesis, diagnosis, detection, quantification, and identification of DNA . It can also be modified and adapted to different types of PCR, such as real-time PCR, reverse-transcriptase PCR, nested PCR, multiplex PCR, long-range PCR, in situ PCR, and assembly PCR .

Limitations

• Primer design: The success and accuracy of PCR depend largely on the design of the primers that are used to initiate the amplification. The primers should be specific, complementary, and optimal in length, temperature, and concentration. Poor primer design can lead to non-specific amplification, primer-dimer formation, or no amplification at all .
• Contamination: PCR is extremely susceptible to contamination by foreign DNA or previously amplified products. Even a trace amount of contaminant DNA can interfere with the amplification and produce false-positive or false-negative results. Therefore, strict precautions and protocols should be followed to prevent and minimize contamination .
• Error-prone: The DNA polymerase enzyme that is used in PCR can introduce errors or mutations in the amplified DNA. This can affect the fidelity and reliability of the PCR products. Some polymerases are more error-prone than others, and the error rate can increase with longer amplicons or more cycles .

• Size limitation: The efficiency and quality of PCR decrease with increasing size of the target DNA region. Longer amplicons require more time, more cycles, more enzyme, and more primers to be amplified. They are also more prone to degradation, inhibition, or mispriming. The maximum size of amplicons that can be reliably amplified by conventional PCR is around 10 kb .

  Precautions using PCR Machine

PCR is a sensitive technique that can amplify very small amounts of DNA from various sources. However, this also makes it prone to contamination, which can lead to false positive results or reduced specificity and sensitivity. Therefore, it is important to follow some precautions when using a PCR machine to ensure the accuracy and reliability of the PCR results. Here are some of the common precautions:

• Designate and use distinct areas for sample preparation, PCR setup, and post-PCR analysis. To avoid contamination from old amplicons, set up the stations on separate benchtops, one for pre-PCR (for PCR reaction setup only) and the other for post-PCR (purifying PCR-amplified DNA, measuring DNA concentration, running agarose gels, and analyzing PCR products). Restrict equipment to these areas.
• Use separate sets of pipettes and pipette tips, lab coats, glove boxes, and waste baskets for the pre-PCR and post-PCR areas. Pipettes and tips are potential sources of contamination, especially if they are reused or shared between different samples or reactions. Lab coats and gloves can also carry DNA from previous experiments or from the environment. Therefore, it is advisable to use disposable pipettes and tips, and change lab coats and gloves frequently. Also, use dedicated glove boxes for handling samples and reagents, and dispose of waste properly.
• Prepare and store reagents for PCR separately and use them solely for their designated purpose. Reagents such as primers, dNTPs, buffer, and polymerase can be contaminated by DNA from previous reactions or from other sources. Therefore, it is recommended to prepare aliquots of reagents in small volumes and store them at appropriate temperatures. Avoid repeated freeze-thaw cycles or exposure to light or heat. Use reagents only for PCR and do not mix them with other reagents.
• Use positive and negative controls in each PCR run. Positive controls are samples that are known to contain the target DNA sequence and can confirm the efficiency and specificity of the PCR reaction. Negative controls are samples that do not contain the target DNA sequence and can detect any contamination or nonspecific amplification in the PCR reaction. Use appropriate positive and negative controls for each PCR run and compare the results with the expected outcomes.
• Clean the cuvette and the hot lid of the PCR machine regularly. The cuvette is the part of the PCR machine that holds the tubes or plates containing the reaction mixtures. The hot lid is the part that presses against the tube or plate lids to prevent evaporation or condensation of the reaction mixtures. Both parts can be contaminated by DNA or reagents from previous runs or from spills. Therefore, it is important to clean them with ethanol or cleaning solution before each run. Also, check that the cuvette holes are clear of dirt or debris that may block the optical path.

• Follow the manufacturer`s instructions for operating and maintaining the PCR machine. The PCR machine is a sophisticated device that requires proper care and maintenance to function optimally. Follow the manufacturer`s instructions for setting up the PCR program, loading the tubes or plates, adjusting the temperature and pressure, running the cycles, reading the results, and troubleshooting any errors or malfunctions. Also, check for any updates or recalls from the manufacturer that may affect the performance or safety of the PCR machine.

  Examples of PCR Machines

There are many types and models of PCR machines available in the market, each with different features and specifications. Here are some examples of PCR machines from different manufacturers:

• Biometra TAdvanced Thermal Cycler Series (Analytik Jena): This series of PCR machines offers 12 different sample blocks, including a high-end 96-well silver block that provides fast heating and cooling rates. The machines also have a high-performance smart lid technology that ensures optimal contact pressure and easy operation. The machines have a large graphical display and a key-pad for easy programming and control. They also have a RAC feature that provides the highest temperature uniformity and reproducibility with zero over- or undershoot of the programmed target temperature.

• MiniAmp™ Plus Thermal Cycler (ThermoFisher Scientific): This PCR machine has a compact size and a 5-inch intuitive color touch-screen that is simple to program and instruct new users. It also has VeriFlex™ Blocks that have three distinct temperature zones that allow precise control of the temperature for PCR optimization. The machine also has WiFi capabilities that enable users to design and securely upload their methods from any mobile device or desktop computer using Thermo Fisher Connect. Additionally, the machine has a front-to-back airflow that allows multiple units to be placed side by side to save valuable bench space.

• PCR Thermal Cyclers (Esco): Esco offers a selection of conventional thermal cycler and real-time thermal cycler models that are designed to meet stringent requirements for all types of PCR processes, including gradient PCR, touchdown PCR, high throughput PCR, in situ PCR, and others that use a range of PCR tubes, strips, plates, and slides. The machines use advanced Peltier temperature control technology to establish and maintain accurate control and fast ramp rates with minimal overshoot or undershoot for process speed and accuracy.

• GET-S SERIES THERMAL CYCLER (Bio-gener): This PCR machine uses an extended service life peltier that has a maximum ramping rate of 4.5 °C/s and a cycle time of over one million. The machine integrates a number of cutting-edge technologies, including the Android operating system, a color capacitive touch screen, multiple block options, an integrated WiFi module, PC software control function, email notification function, a large storage capacity, and more. These features enable the machine to perform PCR with great performance and satisfy more demanding experiment requirements.

• MiniAmp Plus Thermal Cycler and the MiniAmp Thermal Cycler (Delta Science): These PCR machines have a small footprint that fits on any benchtop. They also have VeriFlex™ temperature control technology for PCR optimization and Thermo Fisher Connect for remote access and control. The machines also have a simple user interface with a color touch-screen that is easy to program and instruct new users. The machines can also be used for next-generation sequencing (NGS) library preparation by using the Applied Biosystems™ AmpliSeq™ for Illumina® library prep protocol.

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