The Polymerase Chain Reaction (PCR) is a revolutionary molecular biology technique used to replicate and amplify small amounts of DNA. It has revolutionized the study of genetics, forensics, and evolutionary biology by allowing researchers to quickly and accurately work with large quantities of genetic material from even the smallest samples. The PCR process involves three steps: denaturation, annealing, and extension. Through these steps, multiple copies of a specific sequence of DNA are produced in a relatively short amount of time.
In the denaturation step, doublestranded DNA (dsDNA) is heated up to 9598°C so that it separates into individual strands. This occurs because heating causes hydrogen bonds between base pairs in dsDNA to break apart due to thermal energy input. Once separated into single strands, the two strands can now act as templates for replication during the next step.
In the annealing step, two oligonucleotide primers are used which each bind adjacent regions on opposite sides of an area containing target DNA sequences known as amplicons or amplifiable targets (ATs). These primers provide attachment points for a polymerase enzyme which catalyzes nucleic acid synthesis when temperature cools down to about 50°C60°C . During this stage, complementary bases on both strands form hydrogen bonds until they come together forming what is called hybridization or Annealing. After hybridization takes place between primer and AT sites ,the thermally stable complex allows amplification process begin at lower temperatures than those required for initial strand separation thus conserving additional energy costs associated with repeat denaturation cycles .
Finally inExtension step , once the primers have been bound to their respective single stranded templates by hybridization , Taq polymerase enzyme will facilitate nucleic acid synthesis using deoxyribonucleoside triphosphates as substrate molecules . The heat resistant Taq polymerase enzyme uses template ssDNA strand as guide track while adding corresponding complimentary bases along its length resulting in production new double stranded product that contains one original parent strand plus one newly synthesized daughter strand . As soon as enzyme finishes its job all three components dissociate from each other forming two new intact complementary dsDNAs ready be reused again in upcoming cycle thus providing exponential increase output over time creating millions billions copies identical molecule from original sample within few hours .
This completes one cycle PCR reaction after which same three steps need repeated several times order yield sufficient amount target gene amplified from initial sample size . In conclusion , PCR technology not only offers tremendous potential applications but also provides huge advantages over traditional laboratory techniques such increased sensitivity detection low copy number templates faster turnaround times shorter duration experiments greater accuracy reproducibility when executed correctly under optimal conditions hence proving why it still considered gold standard method detecting analyzing genes today without sacrificing cost effectiveness quality results desired investigator’s field research work
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