DNA polymerase (DNAP) is an enzyme that plays a significant role in DNA replication. DNAP produces nucleotide triphosphates, which are considered the building blocks of DNA. These enzymes work in pairs to create two identical DNA strands from one existing DNA molecule. The job of DNA polymerase is to “read” the existing DNA, which creates two new strands identical to the existing ones.Polymerase was discovered in 1956 by Arthur Kornberg and colleagues while studying Escherichia coli (E. coli). In 1959, he was awarded the Nobel Prize for his discovery. The discovery of DNAP led to the invention of polymerase chain reaction (PCR) in 1983 by Kary Mullis. PCR can assist in extracting DNA for sequencing and cloning, studying gene expression patterns, and the phylogenic DNA analysis from ancient sources, among other applications. Read on to learn more about DNAP, its process, and some equipment needed for your DNA testing laboratory.
DNA Polymerase: What Is the Process?
DNA polymerase must be present every time a cell divides. Before replication, helicase, an enzyme, unravels the DNA, opening the double strands so that two single strands can be used in the replication process. Next, the DNA polymerase adds free nucleotides to the strand being formed. DNA polymerase can only add to a pre-existing group and cannot form its own strand; therefore, a primer is required.
Primers are created by another enzyme called primase and are typically formed from DNA and RNA bases, with the first two bases always being RNA. The process of DNAP is fairly accurate. However, there is a mistake in roughly every one billion pairs. Because of this, DNAP also “proofreads” its work after replication so that any errors can be corrected.
Equipment in a DNA Lab
DNA polymerase is used in many different applications, from DNA cloning to test results. If you’re starting from scratch, you’ll need everything from accessories, such as pipettes, to large pieces of automated laboratory equipment, like liquid handling machines. Some necessary equipment includes:
- DNA isolation kits. Plasmid DNA purification is required to perform processes like sequencing, cloning, and library preparation. Invest in plasmid purification kits to extract high-quality plasmids from fungal and bacterial cell types.
- PCR machines. A PCR machine allows you to conduct various DNA tests simultaneously. While they have many advantages, one is lessening the risk of contamination.
- Restriction enzymes. Using restriction enzymes can help researchers see more closely and are accustomed to look at a single segment of DNA. Restriction enzymes separate the other strands from the original. If a researcher sees a mutation, restriction enzymes also let you see the mutation more closely.
- DNA analyzer. An analyzer detects nucleotide sequences. The sequencing and analysis of DNA fragments are performed using capillary electrophoresis. Capillary electrophoresis causes negatively-charged DNA to migrate, which helps to separate DNA fragments. Essentially, it allows a researcher to match a different sample if necessary.
- Thermocycler. A thermocycler holds tubes that contain a DNA PCR mixture. It raises or lowers the temperature, which splits and amplifies. This allows for DNA replication and multiple copies of the same strand.
- Gel electrophoresis machine. A gel electrophoresis machine takes film of the findings for further study. Precise photos of the gel electrophoresis machine help one see closely.
Depending on your DNA polymerase application, you should invest in further equipment, such as pipettes and pipette tips, centrifuges, hybridization ovens, electrophoresis systems, and more.
Contact Hudson Robotics to discuss options for automating your DNA laboratory to find the lab equipment that perfectly meets your needs.
Sources:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2391090/
https://www.sciencedirect.com/topics/neuroscience/dna-polymerase
https://www.news-medical.net/life-sciences/What-is-DNA-Polymerase.aspx
https://www.nature.com/scitable/definition/dna-polymerase-dnap-1/