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Innovations in Noninvasive Instrumentation and Measurements
Published in Robert B. Northrop, Non-Invasive Instrumentation and Measurement in Medical Diagnosis, 2017
The polymerase chain reaction (PCR) is a molecular biology/biochemical protocol used to exactly replicate (amplify) a single piece of DNA (oligo) from a sample to create thousands to millions of copies of that particular DNA sequence (Riley 2005). These copies allow other analytical techniques to more easily identify the base sequences of the oligo. The PCR allows the DNA from a sample as small as a fraction of a genome, a single-cell nucleus, or a mitochondrion to be characterized in terms of its base sequences. The PCR has many applications, ranging from criminal science to genomics. However, in this section, we will address the PCR applications in medical diagnosis.
Gene Therapy and Gene Correction
Published in Yashwant V. Pathak, Gene Delivery Systems, 2022
Manish P. Patel, Sagar A. Popat, Jayvadan K. Patel
From embryo formation to death, all life has a designated sequence. Any minor changes in any step may lead to problems. It was clinically observed that 6% of newborns were characterized with congenital heart disease, which mainly occurs due to a genetic abnormality (Movafagh et al. 2008). The abnormalities in a genome that can be identified by a change in the DNA sequence of a gene is called a mutation. This results in the development of a faulty protein, which causes genetic problems by changing normal function (Griffiths et al. 1999).
Biomolecules
Published in Volodymyr Ivanov, Environmental Microbiology for Engineers, 2020
Determining the nucleotide sequence in DNA is extremely important because it can be used for identifying microbial strains, detecting strains in an ecosystem, comparing strains, improving strain properties by directional changes in DNA, and carrying out genetic analyses of microbial diversity in ecosystem and of the evolution of life on Earth. The process of determination of DNA sequences is called DNA sequencing.
Gene doping: Present and future
Published in European Journal of Sport Science, 2020
Rebeca Araujo Cantelmo, Alessandra Pereira da Silva, Celso Teixeira Mendes-Junior, Daniel Junqueira Dorta
One of the techniques that is most frequently employed to identify therapeutic vectors and transgenic expression is real-time Polymerase Chain Reaction (PCR), which allows both the identification of extraneous DNA sequences (e.g. vector sequences or copy number variations of a candidate gene), as well as quantitative and qualitative measurements of mRNA transcribed by a candidate gene. Although the PCR methodology is subject to several types of external contamination, which can provide false-positive results, a research group from Australia has been able to circumvent this issue and to establish a real-time PCR-based assay to detect EPO cDNA and plasmid sequences. Moreover, the synthetic reference material developed by Baoutina et al. (2016) can be applied not only in PCR, but also in the advancement of genomics and transcriptomics.
Development of capability for genome-scale CRISPR-Cas9 knockout screens in New Zealand
Published in Journal of the Royal Society of New Zealand, 2018
Francis W. Hunter, Peter Tsai, Purvi M. Kakadia, Stefan K. Bohlander, Cristin G. Print, William R. Wilson
Few New Zealand scientists will be unaware of CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9) as a remarkable new technology that is transforming many aspects of biological research. This ancient apparatus of adaptive immunity, widely distributed in archaea and bacteria (Bhaya et al. 2011), is based on the recognition of foreign nucleic acids rather than peptides, facilitating its adaptation as a molecular biology tool. CRISPR-Cas systems have been reengineered to seek out and manipulate genes in living cells (including higher eukaryotes) providing techniques for precise re-writing of DNA sequences (gene editing) (Komor et al. 2016; Paquet et al. 2016), inactivating genes (gene knockout) (Kleinstiver et al. 2016), transcriptional or epigenetic modification of levels of gene expression (Larson et al. 2013; Perez-Pinera et al. 2013), dynamic imaging of genomic loci and RNA movement in cells (Chen et al. 2013) and as a diagnostic for facile and extremely sensitive detection of specific nucleic acids (Gootenberg et al. 2017). These tools are already finding widespread and diverse application in the biological and biomedical sciences in New Zealand and may yet play an important role in our biosecurity through, for example, CRISPR-mediated gene drive technology (Hammond et al. 2015).