Question

What is the full form of PCR? Explain the use of PCR in biotechnology.

Answer 1

The full form of PCR:

1. PCR stands for a polymerase chain reaction.
2. PCR is a standard molecular biology technique for making numerous copies of a given DNA.
3. Enzymatic replication of DNA serves as the basis for the PCR technique.
4. In PCR, primer-mediated enzymes are used to amplify a short piece of DNA.
5. New DNA strands that are complementary to the template DNA are created by DNA polymerase.
6. The DNA polymerase can add a nucleotide to the pre-existing 3’-OH group only. Therefore, a primer is required.
7. As a result, new nucleotides are added to the 3' prime end of the DNA polymerase.

PCR is based on three simple steps required for any DNA synthesis reaction.

1. Denaturation:

1. In this step, the template is denatured into single strands.
2. When the reaction mixture is heated to 94°C for between 0.5 and 2 minutes, denaturation takes place.
3. This breaks the hydrogen bonds between the two strands of DNA and converts them into a single-stranded DNA.

2. Annealing:

1. Primers are annealed to each original strand to synthesize new DNA strands.
2. For around 20 to 40 seconds, the reaction temperature is decreased to 54 to 60 °C.
3. Here, the primers attach to the template DNA's complementary sequences.

3. Elongation:

1. The temperature is increased to 72-80°C at this stage.
2. The Taq polymerase enzyme adds the nucleotides to the primer's 3' end.
3. Finally, the new DNA strands are extended from the primers.

The use of PCR in biotechnology:

1. PCR is a fast and efficient method for amplifying DNA or RNA sequences from a variety of sources.
2. The final product can be sequenced, examined by gel electrophoresis, or cloned into a plasmid for experimental purposes after PCR.
3. It is a molecular biology technique for multiplying a single copy of DNA into millions of copies.
4. This approach can be used to amplify the gene that is used for vector insertion.
5. For the amplification of the DNA sample received from the investigation site, PCR has often been utilized in DNA fingerprinting methods.
6. In the paternity testing process, DNA samples can be acquired and amplified for detection.
7. This approach aids in the detection of mutations that occur in genetic illnesses.
8. This approach can be used to amplify and study any gene.