Polymerase Chain Reaction (PCR) testing is a molecular biology technique used to amplify and analyze DNA or RNA samples. So, To perform PCR testing, which list of laboratory apparatus and equipment is required?
You need a thermocycler and appropriate thin-walled PCR sample tubes.
PCR tubes , Thermocycler and your components including Master mix ( dNTps, Mgcl2, DMSO,ddH2o, primers), also for sure your Template and the Polymerase enzyme
Hello Al-Haythm Ahmed Al-Essawy
First and foremost, you will need a separate laboratory space/room maintained at the required working temperature to conduct the PCR test. PCR should be conducted in a PCR hood which is a workspace enclosed on three sides that provides a space for performing the amplification of DNA.
For the PCR technique, the most important requirement is the thermal cycler also called PCR machine which is an instrument that amplifies target nucleic acid sequences into millions of copies via PCR. Besides the thermal cycler, you will require a -20-degree freezer to store the samples and PCR reagents.
The main PCR reagents include:
1. The PCR primers which are short strands of nucleotides of approximately15–30 bases. PCR primers are complementary to the template DNA and serve as a DNA synthesis starting point for DNA polymerase.
2. DNA Taq polymerase is a type of DNA polymerase that is active at high temperatures and since PCR is carried at high temperatures, this enzyme is the best choice. This is the molecule that helps in amplifying the sample.
3. Deoxynucleotide triphosphates (dNTPs) are the building blocks of nucleic acids. They consist of four basic nucleotides - dATP, dCTP, dGTP, and dTTP, and they are needed to synthesize DNA by DNA Taq polymerase.
4. PCR buffer which provides a suitable chemical environment for DNA polymerase to perform efficiently. They help maintain a stable pH during PCR and ensure that the reaction is conducted under optimal conditions.
5. Lastly, the DNA template which is the specific sequence of DNA that is the gene of interest which needs to be amplified. It is the starting material for the PCR reaction.
Additionally, you will require a tabletop microcentrifuge to centrifuge the sample/reagents, a vortex mixer to mix small vials of liquid, and a water bath set at 37 degree C (optional).
Among the consumables, you will need 0.2ml PCR tubes, filter pipette tips that prevents PCR contamination, and micropipettes {P2 (0.2 - 2ul), P10(0.5 – 10ul), P20 (2 – 20ul), P50 (5 – 50ul), P100 (10 – 100ul) and P200 (20 – 200ul)}.
Please note that if you wish to analyze RNA samples, Taq polymerase will not work on RNA samples. The incorporation of the enzyme reverse transcriptase (RT), however, can be combined with traditional PCR to allow for the amplification of RNA molecules.
Best.
For equipment, besides the general lab equipment such as pipette, centrifuge, vortex, etc., you need a thermocycler such as this: https://lab.plygenind.com/product/tc1000-s-thermal-cycler-standard
You probably also need an electrophoresis system to analyze the PCR products. For a more comprehensive list, see the following:
https://lab.plygenind.com/supplies-and-equipment-to-run-a-pcr-test-a-basic-overview
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