The principle of the Polymerase chain reaction PCR machine is based on DNA polymerase in vitro replication of specific DNA sequences. This is a way of generating multiple DNA sequences of a specific strain. The DNA strain can be replicated selectively through a large amplification process. The PCR technique enables the purification and cloning of specific DNA. The matrix DNA out of scope of interest will not be varied. The single of the selected DNA is amplified from a combination of other sequences.
Where is PCR applied?
The PCR technique is indispensable in cellular and molecular biology. The “acellular cloning”, in a few hours through automated processing is invaluable compared to the standard processing of molecular cloning that easily takes days to happen. The PCR has proved to be highly useful in diagnostic purposes as it easily identifies the exact DNA sequence of organisms in biological fluids required for the study. It also helps in genetic fingerprinting for judicial inquiries. Botanists and zoologists can identify plant and animal varieties, food quality testing is also done through a PCR. For characterization and conservation of the vast genetic diversity, the sequence of the genome is important for livestock breeds, the measurement of polymorphisms scattered at loci through the genome, the study of large-scale gene transcription at a complex biological scale, and molecular technology with integrated interdisciplinary skills.
How does the PCR work?
A reactive mixture that includes Taq polymerase, the primers, and the four along with an excess solution of deoxyribonucleoside triphosphates (dNTPs) for buffer carry out the polymerase chain reaction.
The tubes are filled with the reactive mixture. They are made to pass through thermal cycler heating blocks of repetitive temperature multiple times.
The enclosed apparatus holding the sample tubes experiences quick temperature variations from 0 to 100°C. A single cycle includes three periods within tens of seconds and the process is further broken into three steps.
Denaturation is where two strands of the DNA are separated with an increase in temperature at 94°C. The DNA matrix is denatured at this temperature. The hydrogen bonds no longer exist when the temperature exceeds 80°C, the double DNA strands are denatured to a single strand.
Hybridization is performed between 40 and 70°C. When the temperature is decreased hydrogen bonds return to their complimentary stands known as primer hybridization. The short strand sequence that is within the aspect of amplification can hybridize easily compared to those with long stands matrix DNA. Hybridization becomes selective and specific at a higher temperature.
Elongation is the third period that occurs at a temperature of 72°C where a combination of complementary strands is bound by Taq polymerase to the primed single-stranded DNAs using deoxyribonucleoside triphosphates in the reaction mixture catalyzing replication. Selective synthesis takes place, synthesized fragments in the previous cycle are matrix after the predominant strain amplifies corresponding to the DNA where the primers hybridize. After 20–40 cycles an analyzable DNA strain is synthesized. The doubling of the interest of a large sequence is extremely rapid and happens within a short duration.
Why invest in iGeneLabserve PCR Machine?
The iGene PCR workstation works with quality HEPA and prefilters. The air filter systems enable work efficiency up to 95%. The work area can be managed easily with a stainless-steel workbench with standard electric fittings and fluorescent lamps. Manual sliding door sashes cover the front part made of acrylic plastic. To know more about iGeneLabserve products, visit the website https://www.igenels.com/.
