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Relevant bibliographies by topics / Denaturing gradient gel electrophoresis

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Academic literature on the topic 'Denaturing gradient gel electrophoresis'

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Published: 4 June 2021

Last updated: 31 January 2023

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Journal articles on the topic "Denaturing gradient gel electrophoresis"

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Hovig, Eivind, Birgitte Smith-Sørensen, Anton Brøgger, and Anne-Lise Børresen. "Constant denaturant gel electrophoresis, a modification of denaturing gradient gel electrophoresis, in mutation detection." Mutation Research Letters 262, no. 1 (January 1991): 63–71. http://dx.doi.org/10.1016/0165-7992(91)90108-g.

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Choe, Myeongeun, Sung-Jun Hong, Jong-Hui Lim, Yunyoung Kwak, Chang-Gi Back, Hee-Young Jung, In-Jung Lee, and Jae-Ho Shin. "Korean Paddy Soil Microbial Community Analysis Method Using Denaturing Gradient Gel Electrophoresis." Journal of Applied Biological Chemistry 56, no. 2 (June 30, 2013): 95–100. http://dx.doi.org/10.3839/jabc.2013.016.

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Cremonesi, Laura, Paola Carrera, Antonella Fumagalli, Sabrina Lucchiari, Elena Cardillo, Maurizio Ferrari, Sabina Carla Righetti, Franco Zunino, Pier Giorgio Righetti, and Cecilia Gelfi. "Validation of Double Gradient Denaturing Gradient Gel Electrophoresis through Multigenic Retrospective Analysis." Clinical Chemistry 45, no. 1 (January 1, 1999): 35–40. http://dx.doi.org/10.1093/clinchem/45.1.35.

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Abstract Among established techniques for the identification of either known or new mutations, denaturing gradient gel electrophoresis (DGGE) is one of the most effective. However, conventional DGGE is affected by major drawbacks that limit its routine application: the different denaturant gradient ranges and migration times required for different DNA fragments. We developed a modified version of DGGE for high-throughput mutational analysis, double gradient DGGE (DG-DGGE), by superimposing a porous gradient over the denaturant gradient, which maintains the zone-sharpening effect even during lengthy analyses. Because of this innovation, DG-DGGE achieves the double goals of retaining full effectiveness in the detection of mutations while allowing identical run time conditions for all fragments analyzed. Here we use retrospective analysis of a large number of well-characterized mutations and polymorphisms, spanning all predicted melting domains and the whole genomic sequence of three different genes—the cystic fibrosis transmembrane conductance regulator (CFTR), the β-globin, and the p53 genes—to demonstrate that DG-DGGE may be applied to the rapid scanning of any sequence variation.

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Adil, Essahale. "Corrective Measures of Denaturing Gradient Gel Electrophoresis Limitations." Journal of Environmental Science and Technology 8, no. 1 (December 15, 2014): 1–12. http://dx.doi.org/10.3923/jest.2015.1.12.

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Radojkovic, Dragica, and Jelena Kušic. "Silver Staining of Denaturing Gradient Gel Electrophoresis Gels." Clinical Chemistry 46, no. 6 (June 1, 2000): 883–84. http://dx.doi.org/10.1093/clinchem/46.6.883.

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Fodde, Riccardo, and Monique Losekoot. "Mutation detection by denaturing gradient gel electrophoresis (DGGE)." Human Mutation 3, no. 2 (1994): 83–94. http://dx.doi.org/10.1002/humu.1380030202.

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Valášková, V., and P. Baldrian. "Denaturing gradient gel electrophoresis as a fingerprinting method for the analysis of soil microbial communities." Plant, Soil and Environment 55, No. 10 (October 21, 2009): 413–23. http://dx.doi.org/10.17221/132/2009-pse.

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In soil microbial ecology, the effects of environmental factors and their gradients, temporal changes or the response to specific experimental treatments of microbial communities can only be effectively analyzed using methods that address the structural differences among whole communities. Fingerprinting methods are the most appropriate technique for this task when multiple samples must be analyzed. Among the methods currently used to compare microbial communities based on nucleic acid sequences, the techniques based on differences in the melting properties of double-stranded molecules, denaturing gradient gel electrophoresis (DGGE) or temperature gradient gel electrophoresis (TGGE), are the most widely used. Their main advantage is that they provide the possibility to further analyze whole sequences contained in fingerprints using molecular methods. In addition to the analysis of microbial communities based on DNA extracted from soils, DGGE/TGGE can also be used for the assessment of the active part of the community based on the analysis of RNA-derived sequences or for the analysis of sequences of functional genes encoding for proteins involved in important soil processes.

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Willame, Raphaël Boutte, Stana Grubisic, Pierre Balthasart, Annick Wilmotte, and Lucien Hoffmann. "Seasonal cyanobacterial dynamics in a mesoeutrophic reservoir: microscopic counts and DGGE (Denaturing Gradient Gel Electrophoresis)." Algological Studies 129 (September 1, 2009): 71–94. http://dx.doi.org/10.1127/1864-1318/2009/0129-0071.

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Nakatsu, C. H. "Soil Microbial Community Analysis Using Denaturing Gradient Gel Electrophoresis." Soil Science Society of America Journal 71, no. 2 (March 2007): 562–71. http://dx.doi.org/10.2136/sssaj2006.0080.

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Shi, L., G. Yan, Y. Fu, L. Ma, A. Penfornis, and D. Faustman. "Human TAP1 polymorphisms detected by denaturing gradient gel electrophoresis." Tissue Antigens 49, no. 4 (April 1997): 421–26. http://dx.doi.org/10.1111/j.1399-0039.1997.tb02772.x.

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Dissertations / Theses on the topic "Denaturing gradient gel electrophoresis"

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Cadiou, Helene. "Evaluation of denaturing gradient gel electrophoresis (DGGE) as a method of mutation detection." Thesis, University College London (University of London), 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.418093.

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Le, Riche Mia. "Lipoprotein X : biochemical predictors and detection by non-denaturing polyacrylamide gradient gel electrophoresis." Thesis, Stellenbosch : Stellenbosch University, 2007. http://hdl.handle.net/10019.1/18218.

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Assignment (MMed)--University of Stellenbosch, 2007.
ENGLISH ABSTRACT: Lipoprotein X (LpX) is an abnormal cholesterol-containing particle that may be present in the serum of subjects with cholestasis, lecithin:cholesterol acyltransferase (LCAT) deficiency and parenteral nutrition. The biochemistry, metabolism, clinical significance and laboratory analysis of LpX is discussed in this study. This laboratory-based project investigated icteric samples received at the Chemical Pathology laboratory, Tygerberg Hospital, for serum predictors of LpX and the use of a modified non-denaturing polyacrylamide gradient gel electrophoresis system in the detection of LpX. The study showed that the non-denaturing polyacrylamide gradient gel electrophoresis system (2-8%) is a useful test in demonstrating LpX in icteric plasma and has potential for a screening test in LCAT deficiency. Serum concentration of conjugated bilirubin, alkaline phosphatase, gamma glutamyltransferase, free cholesterol, phospholipid, free cholesterol: total cholesterol ratio and conjugated bilirubin: total bilirubin ratio are all good predictors of LpX. The ratio of free cholesterol to total cholesterol (FC/TC > 0.6) was the best predictor of LpX. In the setting of obstructive liver disease LpX is seen in 66% of patients if total cholesterol is > 7.5 mmol/L.
AFRIKAANSE OPSOMMING: Lipoproteien X (LpX) is ‘n abnormale cholesterol-bevattende partikel wat teenwoordig mag wees in die serum van persone met cholestase, lesitien:cholesterol asieltransferase (LCAT) gebrek en parenterale voeding. Die biochemie, metabolisme, kliniese belang en laboratorium analise van LpX word bespreek in hierdie werkstuk. Hierdie laboratorium-gebaseerde projek het geelsugtige monsters ondersoek wat ontvang is by die Chemiese Patologie laboratorium, Tygerberg Hospitaal, vir serum voorspellers van LpX en die gebruik van ‘n gemodifiseerde nie-denaturerende polie-akrielamied gradiënt gel elektroforese sisteem in die demonstrasie van LpX. Die bevindinge was dat die nie-denaturerende polie-akrielamied gradient gel elektroforese sisteem (2-8%) is ‘n nuttige toets om LpX te demonstreer in geelsugtige plasma en het potensiaal as ‘n siftingstoets in LCAT gebrek. Serum konsentrasie van gekonjugeerde bilirubien, alkaliese fosfatase, gamma glutamieltransferase, vry cholesterol, fosfolipied, vry cholesterol:totale cholesterol verhouding en gekonjugeerde bilirubien:totale bilirubien verhouding is alles goeie voorspellers van LpX. Die verhouding van vry cholesterol tot totale cholesterol (VC/TC > 0.6) was die beste voorspeller van LpX. In gevalle van obstruktiewe lewersiekte word LpX gesien in 66% van pasiente as die totale cholesterol meer as 7.5 mmol/l is.

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Surridge, Angela Karen Joanna. "Denaturing gradient gel electrophoresis characterisation of microbial communities in polycyclic aromatic hydrocarbon and polychlorinated biphenyl contaminated soil." Thesis, Pretoria : [s.n.], 2007. http://hdl.handle.net/2263/25070.

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Fossil fuels are currently the primary industrial energy source on Earth. They are principally composed of complex hydrocarbons in either long-chain or cyclic conformation. Industrial use of petroleum, diesel, oil, tar and other coal-derived products inevitably leads to pollution of the environment. The most serious pollution is caused by polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) that are not easily removed from soil after a spill. Long-chain and cyclic conformation makes fossil fuel hydrocarbons difficult to break down. However, certain free-living soil microorganisms have adapted to utilising these PAHs/PCBs as a source of energy. In many cases, their efficacy is greatly enhanced by the presence of plants. By inhabiting the rhizosphere, microbes form a mutualistic relationship with the plant, receiving nutrients from it and in return providing a less polluted environment in which the plant can grow. The purpose of this study was to elucidate some of the microbial population diversity in PAH/PCB-polluted soils in South Africa through the use of denaturing gradient gel electrophoresis (DGGE). In an initial study, DGGE was employed to separate soil communities in polluted and unpolluted soils into a genetic fingerprint, the main bands of which were sequenced and subjected to a BLAST analysis through a database for possible identification of species present. Phylogenetic and distance studies indicated that unpolluted soils have a far greater species diversity. It thus was evident that PAH/PCB pollution of soil leads to a decrease in microbial diversity by selecting for microorganisms with the ability to activate metabolic pathways allowing them to utilise the pollutants as an alternative source of carbon. Population diversity of pro- and eukaryotes found within polluted and non-polluted soils was compared. DGGE was employed to determine the genetic fingerprint of each population. Following this, dendogram analyses based on Shannon indices were done to determine PAH breakdown potential of prokaryotic vs. eukaryotic communities. A higher diversity and better adaptation potential were evident within prokaryotic than eukaryotic communities in pollution-stressed environments, indicating that the prokaryotic component of these samples had the greatest PAH-metabolism potential. To determine the capacity for PAH/PCB metabolism by the organisms within the soil samples being studied, the presence of xylE and ndoB genes, responsible for toluene/xylene and naphthalene biodegradation, respectively, was determined. DGGE was performed to analyse genetic diversity between these two genes, based on community fingerprints. Polluted soil communities tended to have comparable community diversity within their functional genes, depending on their physical situation, plant species proximity and soil conditions. In general, soil contained indigenous microbes with a high natural potential for biodegradation of PAHs/PCBs. A portion of the 16S gene of eight bacterial isolates representing the most dominant culturable taxa in the polluted soils was sequenced and analysed for identification purposes. These identifications were conducted in conjunction with the use of the catabolic gene probes xylE and ndoB to establish the hydrocarbon degrading capacity of the isolates. Pseudomonas, from the rhizosphere of Cyperus esculentus, was the most common PAH-degrading genus found in this study. Considering the well-established rhizosphere competence and PAH-degrading capacity of Pseudomonas, this genus seems to be the best suited for bioaugmentation purposes in South Africa. The presence of the nifH gene, the general marker gene of nitrogen-fixing bacteria in communities from unpolluted and polluted soils, was determined. It was hypothesised that bioremediation could be enhanced by nitrogen addition to polluted environments. Nested-PCR of the nifH gene was conducted on a diagnostic basis and was followed by DGGE of the product to determine the functional gene diversity within pollution-dwelling, nitrogen-fixing bacterial communities. Nitrogen-fixing microorganisms were present in all the soils sampled but, in only 80% of the pure cultures isolated from polluted and unpolluted soils and rhizospheres. Although different rhizospheres and pollutants were examined, it was found that of the polluted soils studied, most nifH gene diversity of polluted soils existed within machinery oil polluted, wood chip mulched, non-rhizosphere soil. Thus, it would appear that the more polluted the soil the higher the free microbe nitrogen fixation diversity possibly due to environmental stress.
Thesis (PhD (Microbiology))--University of Pretoria, 2007.
Microbiology and Plant Pathology
unrestricted

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Ellwood-Thompson, Rhianedd Eleri. "Occurrence and transmission of Wolbachia endosymbionts in the oak gall wasp community : application of denaturing gradient gel electrophoresis." Thesis, Cardiff University, 2004. http://orca.cf.ac.uk/55379/.

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Eight Wolbachia variants were identified in the wasp community. Identical Wolbachia variants were identified in inquiline and parasitoid wasp species suggesting that horizontal transmission of Wolbachia occurs in this community

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Strandgren, Charlotte. "Studies of the Diversity of Lactobacillus spp. in Fecal Samples Using PCR and Denaturing Gradient Gel Electrophoresis." Thesis, Linköpings universitet, Institutionen för klinisk och experimentell medicin, 2008. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-15560.

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Allergic diseases, for example asthma and eczema, are nowadays considered belonging to the most common chronic diseases amongst children in the West, but the cause for this increase in allergy prevalence is unknown. Since studies have indicated a connection between children's exposure of microorganisms during infancy and risk of developing allergic disease, it is suggested that this exposure is a crucial factor in question of allergy development or not. Other studies have established differences in microflora composition between healthy children and children with allergic disease, and several studies have shown that probiotic therapy can give positive results in both prevention and treatment of allergic diseases. The aim of this master's thesis was to develop a method, using PCR and denaturing gradient gel electrophoresis, to study the diversity of Lactobacillus spp. in fecal samples retrieved from a study of the probiotic strain L. reuteri ATCC 55730. The developed method was successful in detecting lactobacilli in fecal samples, but three other bacterial genera commonly found in humans were also amplified. Comparison of average numbers of detected bacterial strains and lactobacilli strains between samples belonging to the probiotics and placebo groups, respectively, showed higher numbers for the probiotics group. Also, the only fecal samples that contained L. reuteri belonged to the probiotics group. Although the results are far from statistically significant, they support the theories that probiotics may influence the intestinal microbiota.

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Keyser, Maricel. "PCR detection, denaturing gradient gel electrophoresis (DGGE) fingerprinting and identification of the microbial consortium in different types of UASB granules." Thesis, Link to online version, 2006. http://hdl.handle.net/10019/558.

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Venter, Leandra. "Presence of potentially pathogenic heterotrophic plate count (HPC) bacteria occurring in a drinking water distribution system in the North-West Province, South Africa / by Leandra Venter." Thesis, North-West University, 2010. http://hdl.handle.net/10394/4380.

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There is currently growing concern about the presence of heterotrophic plate count (HPC) bacteria in drinking water. These HPC may have potential pathogenic features, enabling them to cause disease. It is especially alarming amongst individuals with a weakened immune system. South Africa, the country with the highest incidents of HIV positive individuals in the world, mainly uses these counts to assess the quality of drinking water in terms of the number of micro-organisms present in the water. These micro-organisms may be present in the bulk water or as biofilms adhered to the surfaces of a drinking water distribution system. The current study investigated the pathogenic potential of HPC bacteria occurring as biofilms within a drinking water distribution system and determined the possible presence of these micro-organims within the bulk water. Biofilm samples were taken from five sites within a drinking water distribution system. Fifty six bacterial colonies were selected based on morphotypes and isolated for the screening of potential pathogenic features. Haemolysin production was tested for using sheep-blood agar plates. Of the 56, 31 isolates were ?-haemolytic. Among the 31 ?-haemolytic positive isolates 87.1% were positive for lecithinase, 41.9% for proteinase, 19.4% for chondroitinase, 9.7% for DNase and 6.5% for hyaluronidase. All of the ?-haemolytic isolates were resistant to oxytetracycline 30 ?g, trimethoprim 2.5 ?g and penicillin G10 units, 96.8% were resistant to vancomycin 30 ?g and ampicillin 10 ?g, 93.5% to kanamycin 30 ?g, 74.2% to chloramphenicol 30 ?g, 54.8% to ciprofloxacin 5 ?g, 22.6% to streptomycin 300 ?g and 16.1% to erythromycin 15 ?g. Nineteen isolates producing two or more enzymes were subjected to Gram staining. The nineteen isolates were all Gram-positive. These isolates were then identified using the BD BBL CRYSTALTM Gram-positive (GP) identification (ID) system. Isolates were identified as Bacillus cereus, Bacillus licheniformis, Bacillus subtilis, Bacillus megaterium, Bacillus pumilus and Kocuria rosea. 16S rRNA gene sequencing was performed to confirm these results and to obtain identifications for the bacteria not identified with the BD BBL CRYSTALTM GP ID system. Additionally identified bacteria included Bacillus thuringiensis, Arthrobacter oxydans and Exiguobacterium acetylicum. Morphological properties of the different species were studied with transmission electron microscopy (TEM) to confirm sequencing results. All the isolates displayed rod shaped cells with the exception of Arthrobacter oxydans being spherical in the stationary phase of their life cycle. Bulk water samples were taken at two sites in close proximity with the biofilm sampling sites. The DNA was extracted directly from the water samples and the 16S rRNA gene region was amplified. Denaturing gradient gel electrophoresis (DGGE) was performed to confirm the presence of the isolates from the biofilm samples in the bulk water samples. The presence of Bacillus pumilus and Arthrobacter oxydans could be confirmed with DGGE. This study demonstrated the presence of potentially pathogenic HPC bacteria within biofilms in a drinking water distribution system. It also confirmed the probable presence of two of these biofilm based bacteria in the bulk water.
Thesis (M.Sc. (Microbiology))--North-West University, Potchefstroom Campus, 2010.

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Lindelof, Kara L. "Contribution of Biosolids-derived Bioaerosols to the Airborne Microbial Population." University of Toledo / OhioLINK, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1302299544.

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PRUDEN, AMY J. "BIODEGRADATION OF METHYL TERT -BUTYL ETHER." University of Cincinnati / OhioLINK, 2002. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1027943573.

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Van, Niekerk Bertina Freda. "Functional and structural diversity of the microbial communities associated with the use of Fischer–Tropsch GTL Primary Column Bottoms as process cooling water / van Niekerk B.F." Thesis, North-West University, 2011. http://hdl.handle.net/10394/7284.

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Despite emerging water shortages, most water is only used once, and often with low efficiency. However, with appropriate treatment, water can be re–used to reduce the demand on freshwater sources. The Department of Water Affairs, South Africa, promotes industries to reduce discharges into water resources in order to sustain an overall good water quality of all water systems. All of this ultimately leads to industries striving towards zero effluent discharge. Primary Column Bottoms (PCBs) is a wastewater stream derived from the Fischer–Tropsch Gas to Liquid process and consists mainly of organic acids, but no nitrogen or phosphorous, which by implication excludes possible biodegradation. In the operation of cooling towers in industrial processes, cooling water quality has a direct impact on the cooling performance of the system, where nutrient levels may affect fouling, scaling and corrosion observed in the cooling towers. Fouling, scaling and corrosion affect the operating efficiency of cooling water systems and may necessitate the addition of chemical agents to control these phenomena. This has a financial and labour time impact on the operation of these systems. In this study a mini cooling tower test rig was operated with a synthetic PCB effluent as cooling water and various cycles of concentration, pH and linear flow velocities (LFVs). A constant delta temperature of 10 °C was maintained. Cycles of concentration (COC) evaluated included 2, 4 and 6 cycles of concentration and linear flow velocities evaluated was 0.6 m/s, 0.9 m/s and 1.2 m/s. Fouling, scaling and corrosion rates were determined using corrosion coupons and heat exchanger tubes for mild steel and stainless steel. Besides the evaluation of the various operational parameters for fouling, scaling and corrosion, the possibility for chemical oxygen demand (COD) removal by operating the cooling tower as a bioreactor was also evaluated. To this end nutrient correction was applied to the reactor to allow for a CNP ratio of 100:10:1. With regard to fouling, scaling and corrosion, mild steel was more affected by fouling, scaling and corrosion compared to stainless steel where almost no fouling, scaling and corrosion was observed. Overall increased linear flow velocities resulted in higher fouling and scaling rates, whereas lower linear flow velocities resulted in decreased corrosion rates. In terms of cycles of concentration, increased COC resulted in higher fouling, scaling and corrosion rates. Despite the high nutrient removal levels, the accompanying fouling, scaling and corrosion was still below the particular industry’s guidelines. Besides physical–chemical evaluation of the towers under the various operational conditions, culture–dependent and culture–independent methods were also employed. Concerning culture–dependent approaches the study demonstrated that aerobic and anaerobic organisms are present in both the planktonic and sessile phase of the cooling tower reactors. Heterotrophic aerobes were found to be the most abundant under all the operating conditions. Sulphate reducing bacteria were more abundant in the sessile phase of the cooling towers, and the presence of high sulphate levels in the experiments could be indicative of the sulphate reducing bacteria actively participating in the microbial community. Lower than expected corrosion levels, however, suggest that a combination of the organisms in the biofilm rather than sulphate reducing bacteria alone, contributed to the corrosion rates observed. Culture–independent methods, specifically phospholipid fatty acid analysis supported the results from the culture–dependent methods. Furthermore results demonstrated that linear flow velocity had a greater effect on the community structure than cycles of concentration. Finally molecular methods, specifically denaturing gradient gel electrophoresis, found that increasing cycles of concentration resulted in increased microbial community diversity, while increasing linear flow velocity resulted in decreased microbial community diversity. Regarding COD removal, nutrient correction of the synthetic PCB effluent achieved 89.35 % COD removal at 2 COC and 1.2 m/s LFV, while 80.85 % COD removal was achieved at 4 COC at 1.2 m/s LFV. From these results it was recommended that the operation of the cooling tower should be at 4 COC and 1.2 m/s, which despite slightly lower % COD removal, were characterised by fouling, scaling and corrosion rates well within guidelines.
Thesis (M. Environmental Science)--North-West University, Potchefstroom Campus, 2012.

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Book chapters on the topic "Denaturing gradient gel electrophoresis"

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Netzer, Kai-Olaf. "Denaturing Gradient Gel Electrophoresis." In Techniques in Molecular Medicine, 86–95. Berlin, Heidelberg: Springer Berlin Heidelberg, 1999. http://dx.doi.org/10.1007/978-3-642-59811-1_6.

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Roelfsema, Jeroen H., and Dorien J. M. Peters. "Denaturing Gradient Gel Electrophoresis (DGGE)." In Springer Protocols Handbooks, 107–15. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-60327-375-6_8.

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Strathdee, Fiona, and Andrew Free. "Denaturing Gradient Gel Electrophoresis (DGGE)." In Methods in Molecular Biology, 145–57. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-565-1_9.

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Roelfsema, Jeroen H., and Dorien J. M. Peters. "Denaturing Gradient Gel Electrophoresis (DGGE)." In Medical Biomethods Handbook, 79–86. Totowa, NJ: Humana Press, 2005. http://dx.doi.org/10.1385/1-59259-870-6:079.

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Fodde, Riccardo, and Monique Losekoot. "Mutation Analysis by Denaturing Gradient Gel Electrophoresis (DGGE)." In Technologies for Detection of DNA Damage and Mutations, 253–65. Boston, MA: Springer US, 1996. http://dx.doi.org/10.1007/978-1-4899-0301-3_19.

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Knapp, Leslie A. "Single Nucleotide Polymorphism Screening with Denaturing Gradient Gel Electrophoresis." In Methods in Molecular Biology, 137–51. Totowa, NJ: Humana Press, 2009. http://dx.doi.org/10.1007/978-1-60327-411-1_8.

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Green, S. J., M. B. Leigh, and J. D. Neufeld. "Denaturing Gradient Gel Electrophoresis (DGGE) for Microbial Community Analysis." In Handbook of Hydrocarbon and Lipid Microbiology, 4137–58. Berlin, Heidelberg: Springer Berlin Heidelberg, 2010. http://dx.doi.org/10.1007/978-3-540-77587-4_323.

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Green, Stefan J., Mary Beth Leigh, and Josh D. Neufeld. "Denaturing Gradient Gel Electrophoresis (DGGE) for Microbial Community Analysis." In Springer Protocols Handbooks, 77–99. Berlin, Heidelberg: Springer Berlin Heidelberg, 2015. http://dx.doi.org/10.1007/8623_2015_99.

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Lerman, Leonard S. "Electrophoresis of DNA in Denaturing Gradient Gels." In Genetic Engineering, 221–40. Boston, MA: Springer US, 1986. http://dx.doi.org/10.1007/978-1-4615-9456-7_12.

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Jasser, Iwona, Aleksandra Bukowska, Jean-Francois Humbert, Kaisa Haukka, and David P. Fewer. "Analysis of Toxigenic Cyanobacterial Communities through Denaturing Gradient Gel Electrophoresis." In Molecular Tools for the Detection and Quantification of Toxigenic Cyanobacteria, 263–75. Chichester, UK: John Wiley & Sons, Ltd, 2017. http://dx.doi.org/10.1002/9781119332169.ch9.

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Conference papers on the topic "Denaturing gradient gel electrophoresis"

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Apperson, Charles S. "Comparison of microbial communities in monogyne and polygyne Solenopsis invicta colonies by denaturing gradient gel electrophoresis." In 2016 International Congress of Entomology. Entomological Society of America, 2016. http://dx.doi.org/10.1603/ice.2016.112033.

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Arisuryanti, Tuty, Nu-Wei Vivian Wei, and Chris Austin. "Molecular evidence for determination cryptic species of Indonesian swamp eel populations using denaturing gradient gel electrophoresis (DGGE)." In TOWARDS THE SUSTAINABLE USE OF BIODIVERSITY IN A CHANGING ENVIRONMENT: FROM BASIC TO APPLIED RESEARCH: Proceeding of the 4th International Conference on Biological Science. Author(s), 2016. http://dx.doi.org/10.1063/1.4953534.

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Yang, Lijuan, Mengchun Gao, Fangyuan Liang, and Chao Qin. "Microbial Community Analysis During Composting of Digested Sludge and Sawdust by Using Quinone Profile and Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis." In 2009 3rd International Conference on Bioinformatics and Biomedical Engineering (iCBBE). IEEE, 2009. http://dx.doi.org/10.1109/icbbe.2009.5163049.

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Dorfman, Kevin D., and Nabil Laachi. "Band Broadening During High-Throughput Mutation Detection in Microchannels." In ASME 2007 5th International Conference on Nanochannels, Microchannels, and Minichannels. ASMEDC, 2007. http://dx.doi.org/10.1115/icnmm2007-30051.

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Cycling temperature gradient electrophoresis represents a promising method for performing high-throughput DNA mutation detection in a microfluidic platform. Sweeping the temperature between an “all denatured” and “all annealed” state eliminates difficulties introduced by the low thermal mass of the system, while still preserving a mobility difference between the wild type and mutant alleles. We describe a theoretical analysis of this method of mutation detection, based on a multiple-time scales analysis that is valid when the DNA experience many temperature cycles before reaching the detector [1]. We focus on the band-broadening incurred by the interplay between the relaxation time of the chemical system and the thermal oscillations. New results are presented for the case where the denaturing and annealing reactions proceed at identical rates. Our analysis indicates that this separation would be best operated at low electric fields.

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Farache Trajano, Luiza, Rebecca Moore, and Quentin Sattentau. "The Presence of Chemical Cross-Linking Stabilises HIV-1 Envelope Glycoprotein Trimer Antigens in a Model of Intramuscular Immunisation." In Building Bridges in Medical Science 2021. Cambridge Medicine Journal, 2021. http://dx.doi.org/10.7244/cmj.2021.03.001.4.

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Background: The HIV-1 envelope glycoprotein (Env) is the target of antigen design for antibody- based vaccination. In 2019, four trimeric Env vaccines entered an experimental trial: ConM, ConS, and their cross-linked counterparts. The trimers were formulated with MPLA adjuvant. Studies have demonstrated that adjuvants trigger neutrophil infiltration. Neutrophils activate and degranulate releasing proteases, namely elastase and cathepsinG. Aims: To assess the stability and immunogenicity of these vaccines in the presence of adjuvant- recruited neutrophils and their proteolytic enzymes. Methods: Trimers were incubated with commercially-sourced proteases. To analyse stability, samples were reduced, denatured and separated using gel electrophoresis. To assess antibody binding, a trimer-protease incubation was followed by an ELISA. To establish more physiologically relevant conditions, harvested neutrophils were exposed to various adjuvants. The supernatant, shown to contain elastase, was incubated alongside the vaccines. The reducing and denaturing gels, as well as the ELISA, was repeated. Results: Gel analysis revealed that un-crosslinked trimers underwent significant digestion whereas cross-linking conferred enhanced stability. In the presence of neutrophil-sourced protease-containing-supernatant, trimers displayed resistance to digestion. The differential stability profile of Env trimers when exposed to commercially sourced compared to supernatant- derived proteases may be due to the inhibitory effect of human serum on elastase. Antibody epitopes were maintained in vitro. Conclusion: The vaccine antigens are sensitive to enzymatic degradation. This is reduced by cross-linking and human serum.

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Electricwala, A., and T. Atkinson. "TANDEM PURIFICATION OF TISSUE PLASMINOGEN ACTIVATOR BY METAL CHELATE AND AFFINITY CHROMATOGRAPHY." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644831.

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A tandem purification procedure was developed by a combination of metal chelate and affinity chromatography. The conditioned medium from different cell lines producing tissue plasminogen activator was first chromatographed on a zinc chelate-agarose column equilibrated with low ionic strength buffer. After thorough washing, the column was connected to a lysine-agarose column, previously equilibrated with the same buffer. Tissue plasminogen activator was eluted from the zinc chelate column by a gradient of imidazole and the effluent was allowed to flow through lysine-agarose matrix. The two columns were disconnected and after thorough washing, the bound enzyme from lysine-agarose column was eluted with a linear gradient of potassium thiocyanate in the equilibration buffer. This method resulted in a purification factor which varied between 40 to 110 fold. The purity of the isolated enzyme was assessed by sodium dodecyl sulphate-gel electrophoresis and fibrin zymography. The chromatographic procedure described provides a novel method for the rapid purification of tissue plasminogen activator to a high degree of purity.

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Kempfer, A. C., N. Maugeri, C. Farías, E. Bermejo, M. Gimeno, and M. Lazzari. "PURIFICATION AND PARTIAL CHARACTERIZATION OF A BIOACTIVE SUBSTANCE FROM RAT'S VESSEL WALL INDEPENDENT OF PROSTACYCLIN PRODUCTION." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643362.

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Our previous observations provided evidence that a bioactive substance (BAS) with inhibitory effect on platelet aggregation and with inotropic activity on smooth muscle preparations is present in aortic wall of rats treated previously with indomethacin. The ability to inhibit platelet aggregation was used for monitoring its purification and partial characterization. The original sample was extracted by rinsing aortic rings (1.5 mg dried rings) in buffer Krebs (300 μl) for 40 minutes at room temperature. The substance was purified by gel filtration (Bio-gel P-30, Sephadex G-100, Sephadex G-75) and ion exchange chromagra-phy (DEAE cellulose). In the last method, salt gradient elution was performed. Further purification by Sephadex G-50 resulted in removal of 90% of the contaminating substances without a loss of inhibitory activity. The main peak of both chromatographic procedures was analyzed on SDS-PAGE and PAGE with denaturing solvents. The substance was evident by Coomassie Brilliant Blue and periodic acid Schiff staining.In order to determine if the BAS was susceptible to proteolysis, an aliquot of the original sample (29 ug total protein) was incubated with trypsin (final concentration 0.3 μg/ml) and with chymotrypsin (final concentration 3 μg/ml). The BAS activity was not detected. An aliquot of the same original sample was incubated with neuraminidase (final concentration 1.2 units). The BAS activity was detected.The substance appeared to be stable for at least 18 hours at room temperature and 2 hours at 37°C, In addition it was stable over a pH range between 6.8 to 8.6, showing an anionic behaviour.The protein concentration of this substance determined by the method of Lowry was 1 μg/mlPartial characterization supports the conclusion that the substance present in aortic wall of rats is a homogeneous protein, which has a molecular size estimated at 55-65 kDa.

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Heeb, M. J., D. F. Mosher, and J. H. Griffin. "PROTEIN C IS ACTIVATED AND GIVES 110,000 MW COMPLEXES AND PROTEIN S IS CLEAVED AND DECREASED _IN VIVO IN PATIENTS WITH INTRAVASCULAR COAGULATION (DIC)." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644696.

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Immunoblotting studies using denaturing and nondenaturing polyacrylamide gel electrophoresis conditions were performed on 100 plasmas from 88 patients with suspected DIC, in order to determine whether the anticoagulant regulatory proteins C and S (PC and PS) cure altered in vivo during DIC. 70 of these plasmas from 65 patients contained 5-35% of PC antigen in the form of activated protein C (APC) complexed with inhibitor(s).24 normal plasmas showed no detectable APC-inhibitor complexes.The complexes in DIC plasmas had a MW of 110 K on SDS-PAGE, as did complexes formed when APC was incubated with plasma immunodepleted of PC, or when PC in normal plasma was activated with Protac C. On nondenaturing gels, the complex present in 69 of 70 patient plasmas had the same mobility as one of two major bands of complexed APC observed in Protac C-activated normal plasma. One patient plasma contained two forms of PC antigen complex. This patient had suffered a perforated uterus during an abortion. After Protac C activation of the patient plasmas, two APC complexed bands were seen. The 16 patients with >15% complexed PC antigen included 3 with severe infection, 5 with solid tumors, 3 with leukemias, 2 with vascular disease and 3 with other diagnoses. These patients had a higher mortality (69%) than the group as a whole and higher levels of fibrin degradation products. 13 of these 16 plasmas and 56 of the entire group of 100 contained a higher than normal proportion of PS in a cleaved form with an apparent molecular weight lower than intact PS on reduced SDS-PAGE. Mean levels of PS antigen determined by electroimmunoassay for 95 of the plasmas were as follows: entire group, 86% (92%); patients with infection (n=34), 76% (81%); patients with malignancy (n=37), 102% (105%); all others (n=24), 70% (74%) wherfe numbers in parentheses exclude patients with liver disease and 100% normal pooled plasma. These studies suggest that PS is cleaved and decreased in vivo and that PC is activated in vivo and complexed with a 50K MW inhibitor(s) during DIC.

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Jørgensen, M. "HEPARIN INDEPENDENT PURIFICATION OF ANTITHROMBIN III (AT III) BY IMMUNO-AFFINITY CHROMATOGRAPHY RESULTING IN A FUNCTIONALLY INTACT MOLECULE." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643682.

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Previous methods for purification of AT III are based on its heparin-binding capacity. However, in congenital AT III deficiency abnormal inhibitor molecules with impaired binding of heparin and/or thrombin has been reported. The aim of the present study was to develop a purification method based on immuno-affinity chromatography, and thus independent of the heparin binding capacity.Rabbits were immunized with human AT III purified by a three-step procedure involving dextran sulphate precipitation, affinity chromatography on heparin-Sepharose and ion-exchange chromatography on DEAE-Sephadex A-50. Rabbit immunoglobulins against human AT III were isolated by affinity chromatography using purified human AT III coupled to CNBr-activated Sepharose 4B. Trace amounts of immunoglobulin against human albumin, IgG and IgM were removed by solid phase immunoadsorption. The highly purified immunoglobulins against human AT III were coupled to CNBr-activated Sepharose 4B. This anti-AT III-Sepharose was used for single-step purification of AT III from plasma. Elution was performed by Na-citrate buffer at pH 3.0 and the eluted fractions immediately neutralized. The recovery was more than 60%.The purified AT III appeared as a single protein band in SDS-poly-acrylamide gel electrophoresis with or without reduction. Affinity purified AT III and AT III purified by the three-step procedure were indistinguishable when analyzed by crossed immunoelectrophoresis in the absence and the presence of heparin isoelectrical focusing in polyacrylamid gel at a pH 4-6.5 gradient, and SDS-polyacrylamide gel electrophoresis. AT III antigen concentration was determined by electroimmunoassay and the reactive site concentration determined by titration with purified human thrombin using Phe-Pip-Arg-Nan (S-2238) as substrate. The ratio (active site conc.)/(antigen conc.) was identical in the two AT III preparations. It is concluded that this single-step immuno-affinity chromatography gives a high recovery from plasma of a highly purified functionally intact AT III molecule. The purification method is independent of the heparin binding capacity of AT III. This is of particular importance for the purification and characterization of abnormal AT III molecules with impaired heparin-binding site.

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Meijers, Joost C. M., Pim N. M. Tijburg, and Bonno N. Bouma. "INHIBITION OF HUMAN BLOOD COAGULATION FACTOR Xa BY α2-MACROGLO-BULIN." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643838.

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The inactivation of activated factor X (factor Xa) by α2 macroglobulin (α2M) was studied. Irreversible inhibition was observed with the initial formation of a reversible enzyme-inhibitor complex The secopd-order rate constant for the reaction was 8.4 × 104 M−1 min−1. The binding ratio was found to be 2 mol factor Xa/ mol α2M. Interaction of factor Xa with α2M resulted in the appearance of four thiolgroups/molecule α2. The apparent second-order rate constants for the appearance of thiolgroups were dependent on the factor Xa concentration. Sodium dodecyl sulphate gradient polyacrylamide gel electrophoresis was used to study complex formation between α2M and factor Xa. Under non-reducing conditions four factor Xa - α2M complexes were observed. Reduction of these complexes showed the formation of two new bands. One complex (Mr 225000) consisted of the heavy chain of the factor Xa molecule covalently bound to a subunit of α2M, while the second complex (Mr A00000) consisted of the heavy chain of factor Xa molecule and two subunits of α2M. Factor Xa was able to form a bridge between two subunits or α2M, either within one molecule of α2M, or by linking two molecules of The role of the light chain of factor Xa in this process remains to be elucidated. For this purpose, monoclonal antibodies specific for the light chain of factor Xa were prepared. Sodium dodecyl sulphate agarose electrophoresis studies showed that complexes involving more than two molecules of α2M were not formed.

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