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Genomics and Bionanotechnology
Published in Anil Kumar Anal, Bionanotechnology, 2018
PCR can be applied for the quantification of low level of gene expression by utilizing RNA as template. Reverse transcription-PCR (RT-PCR) is the modification of PCR where single-stranded RNA is used as initial template, and enzyme (reverse transcriptase) converts target RNA into complementary DNA (cDNA) copy, which is amplified further by standard PCR method. This approach can be used to amplify messenger RNA (mRNA) or ribosomal RNA (rRNA) rather than DNA, thus detecting specific expression of certain genes during the state of infection. RT-PCR method is therefore useful to detect infection by detecting cDNA of mRNA encoded by a pathogen (Louie et al. 2000).
Recombinant DNA technology
Published in Firdos Alam Khan, Biotechnology Fundamentals, 2018
A cDNA is the copy or complementary DNA produced by (usually) using mRNA as a template. In fact, any RNA molecule can be used to produce cDNA. The DNA copy of an RNA molecule is produced by the enzyme reverse transcriptase (RNA-dependent DNA polymerase; discovered by Temin and Baltimore in 1970) generally obtained from avian myeloblastosis virus.
Glossary of scientific and technical terms in bioengineering and biological engineering
Published in Megh R. Goyal, Scientific and Technical Terms in Bioengineering and Biological Engineering, 2018
cDNA (complementary DNA) refers to the double-stranded DNA complement of an mRNA sequence; synthesized in vitro from a mature RNA template using reverse transcriptase and DNA polymerase (to create the double-stranded DNA).
Artemisia vulgaris essential oil nanoemulsions (AVEO-NE), a novel anti-angiogenic agent and safe apoptosis inducer in MCF-7 human cancer cells
Published in Inorganic and Nano-Metal Chemistry, 2022
Mahjoubeh Irani, Masoud Homayouni Tabrizi, Touran Ardalan, Toktam Nosrat
The expression of genes similar to the research conducted by Khatamian et al. was examined by qPCR method.[25] First, the MCF7 cells were seeded in a 12.5 cm2 culture flask for 24 h and then treated with various doses of AVEO-NE (1, 2, and 4 µg/mL). After 48 h, the RNA content of cells in each group was extracted using the RNeasy mini kit (Qiagen, Hilden, Germany). Then RNA was used as a template for CDNA synthesis and complementary DNA was synthesized using Quantitect Reverse Transcription kit (Qiagen, Hilden, Germany). The target gene primer sets for Cas-9,[26] CAT, SOD, and VEGF[25] are given in Table 1. The GAPDH gene[27] was defined as an internal control gene (Table 1). In order to perform the reaction, a mixture with a volume of 20 μL including 10 μL of Syber Green, 2 μL of specific primer, 1 μL of CDNA, and 7 μL of DW was prepared and analyzed by CFX-96 Biorad.
Assays and enumeration of bioaerosols-traditional approaches to modern practices
Published in Aerosol Science and Technology, 2020
Maria D. King, Ronald E. Lacey, Hyoungmook Pak, Andrew Fearing, Gabriela Ramos, Tatiana Baig, Brooke Smith, Alexandra Koustova
Although PCR is a common method for amplifying DNA, it is also applicable for RNA templates by using reverse transcriptase to synthesize complementary DNA (cDNA) (Saint-Marcoux et al. 2015). Reverse transcription is performed by adding a primer or random hexamer primers and transcriptase before amplification (Lauterbach et al. 2018). DNA is less likely to degrade than RNA samples (Bustin et al. 2009). A small volume of the prepared cDNA sample can be used for further amplification (Saint-Marcoux et al. 2015).
Microbiology in Water-Miscible Metalworking Fluids
Published in Tribology Transactions, 2020
Frederick J. Passman, Peter Küenzi
Reverse transcriptase PCR allows the amplification of RNA that, subsequently, is reversibly transcribed into complementary DNA (cDNA), made possible by the enzyme reverse transcriptase. cDNA is then used as template for any of the PCR methods available.