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Methods of Identifying Microbiological Hazards in Indoor Environments
Published in Rafał L. Górny, Microbiological Corrosion of Buildings, 2020
The terminal restriction fragment length polymorphism (T-RFLP) is an electrophoretic analysis of fluorescently labelled PCR 16S rRNA products, preceded by their digestion with an adequately selected restriction enzyme, crossing the double strand of DNA between nucleotides within a characteristic, several-nucleotide sequence (usually 6–8 bs). The only fragments separated here are those in which in the course of evolution were revealed such mutations which have resulted in the appearance of new particular enzyme-restricted zones. The restrictive fragments profile obtained reflects the relative diversity of microorganisms in the test material. The T-RFLP method is used to analyse populations originating from different environments, including those present in the air [Diel et al. 2005; Lee et al. 2010].
Metagenomics
Published in Ram Chandra, R.C. Sobti, Microbes for Sustainable Development and Bioremediation, 2019
Gaurav Saxena, Narendra Kumar, Nandkishor More, Ram Naresh Bharagava
Molecular techniques are primarily used for the monitoring of microbial communities and to determine the efficacy of bioremediation processes. A number of culturable and nonculturable molecular techniques are currently applied to determine the microbial community composition and structure at contaminated sites. The culture-dependent molecular techniques include various molecular methods, such as amplified ribosomal DNA restriction analysis (ARDRA), randomly amplified polymorphic DNA analysis (RAPD), amplified fragment length polymorphisms (AFLP), length heterogeneity PCR (LH-PCR), single-strand conformation polymorphism (SSCP), automated ribosomal intergenic spacer analysis (ARISA), denaturing gradient gel electrophoresis (DGGE)/temperature gradient gel electrophoresis (TGGE), and terminal-restriction fragment length polymorphism (T-RFLP) (Desai et al. 2011). A short description of conventional molecular techniques with their merits and demerits is presented in Table 10.1. However, a brief description of some of the conventional molecular techniques is provided below:
Recent Developments in the Treatment of Petroleum Hydrocarbon and Oily Sludge from the Petroleum Industry
Published in Sunil Kumar, Zengqiang Zhang, Mukesh Kumar Awasthi, Ronghua Li, Biological Processing of Solid Waste, 2019
Surendra Sarsaiya, Sanjeev Kumar Awasthi, Archana Jain, Saket Mishra, Qi Jia, Fuxing Shu, Jiao Li, Yumin Duan, Ranjan Singh, Mukesh Kumar Awasthi, Jingshan Shi, Jishuang Chen
Molecular methods are important tools for learning the molecular potential of microbes (Liu et al. 2016). Current information about microbial groups is the result of the use of molecular methods (Liu et al. 2010; Yang et al. 2016; Li et al. 2016). The 16S rRNA sequence, reverse genome probing (RGP), restriction fragment length polymorphism (RFLP), oligo-nucleotide matrix array hybridization (OMAH), and denaturing gradient gel electrophoresis (DGGE) are universally accepted techniques for the molecular characterization of microbes’ diversity isolated from the hydrocarbon contamination sites (Al-Hawash et al. 2018). Polymerase chain reaction–based fingerprinting method is less effort-concentrated method that has been widely used to examine microbial diversity. The examination of 16S rDNA has improved from the samples discovered beforehand unidentified diversity in a range of territories (Li et al. 2010; Peixoto et al. 2011; Feng et al. 2011; Zhou et al. 2013; Yang et al. 2016; Liu et al. 2016). The study of diversity has been examined for the explanation of petroleum hydrocarbon microbial diversity (Tardy-Jacquenod et al. 1996; Sette et al. 2007).
Enhanced degradation of phenol in floating treatment wetlands by plant-bacterial synergism
Published in International Journal of Phytoremediation, 2018
Hamna Saleem, Khadeeja Rehman, Muhammad Arslan, Muhammad Afzal
At the end of the experiment, population of total and inoculated bacteria was determined in water, on rhizoplane, and root and shoot interior of T. domingensis. Briefly, 100 µl of treated water and slurry suspension (serial dilutions up to 10−6) of plant material (after surface sterilization) were plated onto the M9 plates containing phenol as a sole carbon source and subsequently placed at 37 °C for incubation. The colony forming units (CFU) were counted after 48 h. A statistically significant number of colonies were picked and subjected to restriction fragment length polymorphism (RFLP) analysis. RFLP reaction was placed at 37 °C for 3 h, where each 1X reaction constituted 7 µL polymerase chain reaction (PCR) product, 1 µL HindIII enzyme, 1.5 µL R-buffer, and 5.5 µL deionized water, to make a total of 15 µL reaction. Restriction fragment length polymorphism (RFLP) product was confirmed by deoxyribonucleic acid (DNA) electrophoresis on 2% agarose gel.