Polymerase chain reaction (PCR) is a technique used to amplify specific regions of DNA. It involves making many copies of a specific DNA fragment using enzymes called polymerases. The process involves heating and cooling a mixture of DNA template, primers, a thermostable DNA polymerase (such as Taq polymerase), and the four deoxynucleoside triphosphates (dNTPs) required for DNA synthesis.
The basic reagents used in PCR are:
- Template DNA: This is the target DNA that you want to amplify. The template DNA can be from any source, including bacteria, viruses, plant or animal cells, or even forensic samples.
- Primers: These are short stretches of DNA that are complementary to the ends of the region of the template DNA that needs to be amplified. Primers typically range in length from 18 to 30 base pairs.
- Taq polymerase: This is a thermostable DNA polymerase that can withstand the high temperatures used in PCR. Taq polymerase is isolated from the bacterium Thermus aquaticus.
- dNTPs: dNTPs are the building blocks of DNA and include deoxyadenosine triphosphate (dATP), deoxycytidine triphosphate (dCTP), deoxyguanosine triphosphate (dGTP), and deoxythymidine triphosphate (dTTP).
- Buffers: PCR requires a buffer that provides a suitable pH, salt concentration, and cofactors that are required for the activity of Taq polymerase.
- MgCl2: Magnesium chloride is also typically added to the reaction mix as a cofactor for Taq polymerase.
There are also different types of PCR, including Real-time PCR, which include the use of fluorescent dyes (probe or primers) or TaqMan probe and a thermal cycler with a fluorescent detector.
It’s important to always keep in mind that to get the best performance out of PCR reactions, it’s important to have high-quality reagents and use them under controlled conditions. We at Pipette.com ensure that we support your science with our products and help you to achieve best results