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Genes and Genomics
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2020
Almost all PCR applications employ a heat-stable DNA polymerase, such as Taq polymerase, an enzyme originally isolated from the bacterium Thermus aquaticus. This DNA polymerase enzymatically assembles a new DNA strand from nucleotides by using single-stranded DNA as a template and DNA oligonucleotides (also called DNA primers), which are required for initiation of DNA synthesis. The vast majority of PCR methods use thermal cycling, that is, alternately heating and cooling the PCR sample in a defined series of temperature steps. These thermal cycling steps are necessary to physically separate the two strands in a DNA double helix at a high temperature in a process called DNA melting. At a lower temperature, each strand is then used as the template in DNA synthesis by the DNA polymerase to selectively amplify the target DNA. The selectivity of the PCR results from the use of primers that are complementary to the DNA region targeted for amplification under specific thermal cycling conditions (Figure 2.17).
Genes and genomics
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2018
Almost all PCR applications employ a heat-stable DNA polymerase, such as Tag polymerase, an enzyme originally isolated from the bacterium Therm.us aquaticus. This DNA polymerase enzymatically assembles a new DNA strand from nucleotides by using ssDNA as a template and DNA oligonucleotides (also called DNA primers) that are required for initiation of DNA synthesis. The vast majority of PCR methods use thermal cycling, that is, alternately heating and cooling the PCR sample in a defined series of temperature steps. These thermal cycling steps are necessary to physically separate the two strands in a DNA double helix at a high temperature in a process called DNA melting. At a lower temperature, each strand is then used as the template in DNA synthesis by the DNA polymerase to selectively amplify the target DNA. The selectivity of PCR results from the use of primers that are complementary to the DNA region targeted for amplification under specific thermal cycling conditions.
Designing biological circuits
Published in Karthik Raman, An Introduction to Computational Systems Biology, 2021
As DNA synthesis technologies become increasingly cheaper and effective, and the reliance on traditional molecular cloning continues to diminish, synthetic biology will enter a log phase of even rapider growth. Nevertheless, the ability to predictively design biological circuits and systems will be limited by our ability to model circuit behaviour and iteratively improve the models based on experimental datasets. For greater success in synthetic biology approaches, it is important to be able to reliably compose parts into larger circuits—and handle the ensuing cross-talk with the existing complex cellular circuitry as well.
Biosafety and biosecurity in Synthetic Biology: A review
Published in Critical Reviews in Environmental Science and Technology, 2019
Lucía Gómez-Tatay, José M. Hernández-Andreu
This subfield of Synthetic Biology involves “the creation of organisms with a chemically synthesized (minimal) genome. This branch of synthetic biology has been made possible by the constant improvements in DNA-synthesis technology over the past years, which now allows the generation of DNA molecules in the range of thousands of base pairs at a competitive price. The aim is to merge these molecules into full genomes and transplant them into living cells, thereby replacing the genome of the host cell and reprograming its metabolism to undertake new tasks” (Deplazes, 2009).