PCR, or polymerase chain reaction, is a technique that allows you to amplify a specific segment of DNA or RNA from a sample. PCR can be used for various purposes, such as diagnosing diseases, identifying pathogens, detecting mutations, analyzing gene expression, and more. But what do you need to run a PCR test in your lab? In this blog post, we will give you an overview of the basic supplies and equipment that are required for PCR.
Supplies
The main supplies that you need for PCR are:
- Nucleic acid template: This is the sample that contains the DNA or RNA sequence that you want to amplify. The template can be extracted from various sources, such as blood, saliva, tissue, cells, etc. Depending on the type of nucleic acid (DNA or RNA), you may need different extraction methods and kits12.
- Primers: These are short synthetic DNA sequences that are complementary to the ends of the target sequence. Primers serve as the starting point for the polymerase enzyme to copy the template. You need two primers for each target sequence, one for each strand. Primers can be designed and ordered from various companies3.
- Nucleotides: These are the building blocks of DNA and RNA. They are composed of a nitrogenous base (A, T, C, G for DNA; A, U, C, G for RNA), a sugar (deoxyribose for DNA; ribose for RNA), and a phosphate group. Nucleotides are added by the polymerase enzyme to extend the primers and synthesize new strands of DNA or RNA. Nucleotides are usually supplied as dNTPs (deoxynucleoside triphosphates) for DNA synthesis or NTPs (nucleoside triphosphates) for RNA synthesis3.
- Polymerase enzyme: This is the protein that catalyzes the PCR reaction. It adds nucleotides to the primers and copies the template. There are different types of polymerase enzymes available for PCR, depending on the application and the desired properties. Some common examples are Taq polymerase, which is derived from a thermophilic bacterium and can withstand high temperatures; Pfu polymerase, which has high fidelity and accuracy; and reverse transcriptase, which can convert RNA into DNA3.
- Buffer: This is the solution that provides the optimal conditions for the PCR reaction. It contains salts, pH stabilizers, magnesium ions, and other additives that affect the activity and specificity of the polymerase enzyme3.
- Master mix: This is a premixed solution that contains all or most of the components needed for PCR, such as nucleotides, polymerase enzyme, buffer, and sometimes primers. Master mixes can simplify the preparation of PCR reactions and reduce pipetting errors34.
- Plasticwares for PCR such as tubes, plates, etc.
Equipment
The main equipment that you need for PCR are:
- Thermal cycler: This is the device that controls the temperature cycles of the PCR reaction. It consists of a metal block with holes where you can insert tubes or plates containing your PCR reactions. The thermal cycler can rapidly change the temperature of the block according to a programmed protocol. The typical protocol consists of three steps: denaturation, annealing, and extension1. These steps are repeated for a number of cycles (usually 25-40) to amplify the target sequence exponentially.
- Centrifuge: This is a device that spins tubes or plates at high speed to separate liquids by density. You may need a centrifuge to spin down your samples before or after extraction, to remove air bubbles from your PCR reactions, or to collect your amplified products after PCR15.
- Vortex mixer: This is a device that vibrates tubes or plates at high speed to mix liquids thoroughly. You may need a vortex mixer to resuspend your samples after extraction, to mix your PCR components before adding them to your reactions, or to dissolve your amplified products after PCR15.
- Pipettes: These are instruments that allow you to transfer precise volumes of liquids from one container to another. You need pipettes to measure and add your samples, primers, nucleotides, polymerase enzyme, buffer, master mix, and other components to your PCR reactions15. You also need pipette tips that fit your pipettes and prevent cross-contamination between your samples and reagents.
- Fridge and freezer: These are appliances that store your samples and reagents at low temperatures to preserve their quality and stability. You may need a fridge to store your samples before or after extraction, and a freezer to store your primers, nucleotides, polymerase enzyme, buffer, master mix, and other reagents15.
- Analysis instruments: These are devices that allow you to visualize and quantify your PCR products. Depending on the type of PCR (qualitative or quantitative) and the detection method (end-point or real-time), you may need different analysis instruments, such as electrophoresis systems, gel documentation systems, spectrophotometers, fluorometers, or qPCR machines34.
Conclusion
PCR is a powerful technique that can amplify a specific segment of DNA or RNA from a sample. To run a PCR test, you need various supplies and equipment that provide the necessary components and conditions for the PCR reaction. In this blog post, we have given you an overview of the basic supplies and equipment that are required for PCR. However, depending on your specific application and protocol, you may need additional or different supplies and equipment. Therefore, you should always consult a professional before performing any PCR experiments.
