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Extremophilic Microbes and their Extremozymes for Industry and Allied Sectors
Published in Ajar Nath Yadav, Ali Asghar Rastegari, Neelam Yadav, Microbiomes of Extreme Environments, 2021
Hiran Kanti Santra, Debdulal Banerjee
A total number of 325 bacterial isolates were obtained from soil and water samples of those regions using 10 different nutrient compositions and characterized using 16S rDNA amplified ribosomal DNA restriction analysis with three restriction endonucleases AluI (from Arthrobacter luteus), MspI (from Moraxella sp.), and HaeIII (from Haemophilus influenzae) leading to the formation of 23–40 groups for different sites having 75% similarity index and adding up-to 175 groups. 16S rRNA gene sequencing identified 175 bacteria belonging to four phyla Actinobacteria (16%), Bacteroidetes (3%), Firmicutes (54%), and Proteobacteria (28%) including 9 different genera with 57 different species.
Carbon Nanotube–Based Electrochemical Biosensors
Published in Li Jun, Wu Nianqiang, Biosensors Based on Nanomaterials and Nanodevices, 2017
Figure 11.15d shows the electrophoresis results of a DNA molecular weight standard (ΦX174RFDNA-HaeIII digest) and the two PCR amplicons, respectively. By shaking the sample in each of the solutions at 40°C for 15 min, the nonspecific binding is removed through stringent washing in three steps using 2 × SSC/0.1%SDS, 1 × SSC, and 0.1% SSC, respectively. Ru(bpy)32+ mediators were used to efficiently transport electrons from the guanine bases to the MWNT nanoelectrode and to provide an amplified guanine oxidation signal as long as target DNA molecules are within the three-dimensional diffusion layer.
First Characterization of PAH-degrading bacteria from Río de la Plata and high-resolution melting: an encouraging step toward bioremediation
Published in Environmental Technology, 2019
Silvina A. Izzo, Silvina Quintana, Mariela Espinosa, Paola A. Babay, Silvia R. Peressutti
The isolates were screened by restriction fragment length polymorphism (RFLP) analysis. PCR products were digested with 5 U of the restriction endonuclease HaeIII (Promega, Madison, WI) and resultant restriction fragments were analyzed by electrophoresis on 2% agarose gel and visualized under UV, by staining with ethidium bromide. The representative isolates of each RFLP pattern were identified by 16S rDNA gene sequencing. 16S rDNA-PCR products were purified with DNA PuriPrep-GP Kit (Inbio-Highway, Tandil, Argentina) and sequenced commercially at INTA (Castelar, Argentina) by using the primers F63 (5′-CAGGCCTAACACAT GCAAGTC-3′) and F530 (5′-GTGCCAGCMGCCGCGG-3′). Both retrieved nucleotide sequences were edited using the FinchTV 1.4.0 program (Geopiza Inc.) and overlapped with the BioEdit software [26]. Each obtained consensus sequence was screened against EzTaxon data base [27] using the BLASTn tool [28]. Sequences were analyzed phylogenetically with the MEGA 5.2 program [29]. Phylogenetic trees were constructed through the neighbor-joining (NJ) algorithm, from a distance matrix calculated following Kimura’s two-parameter model. Stability among the clades was assessed with the 1000-replication bootstrap analysis. Sequences were deposited at the GenBank database under accession numbers KY020297 to KY020338.