National Repository of Grey Literature 76 records found  beginprevious65 - 74next  jump to record: Search took 0.01 seconds. 
Separation and purification of nucleic acids with the use of magnetic microparticles
Vlachová, Jana ; Rittich, Bohuslav (referee) ; Španová, Alena (advisor)
Application of magnetic microparticles for nucleic acid isolation from real samples is a modern and fast separation procedure. This method is suitable for application in laboratory practice. The aim of the bachelor work was isolation of DNA from a dairy product (Activia sweet white) in the PCR-ready quality. DNA isolation was carried out by magnetic microparticles P(HEMA co GMA) in the presence of 2 M NaCl and 16% PEG 6000 or 8% PEG 6000. According to the declaration of the producer the presence of DNA from probiotic bacteria of the genera Lactobacillus and Bifidobacterium was confirmed by PCR.
Application of real-time PCR in microbiological analysis of food-stuffs
Novotná, Eva ; Michaela, Kotianová (referee) ; Španová, Alena (advisor)
Probiotic lactic acid bacteria of genus Lactobacillus play an important role in the digestive tract of human, e.g. protect against pathogenic microorganisms. Lactobacilli are often present in various food products. Lactic acid bacteria of the genus Lactobacillus can be detected by polymerace chain reaction (PCR) using specific primers LbLMA1 and R16. The bacterial DNA was isolated from Actimel Natur probiotic product by the phenol extraction method. DNA was amplified using real time PCR. Specific PCR products were detected using fluorescent intercalaction dye SybrGreen. The specific PCR products were verified by melting curve analysis (Tm 85°C) and by agarose gel electrophoresis (PCR products of 250 bp was amplified).
DNA analysis of nonpathogenic clostridia isolated from cheeses
Chroboková, Maria ; Trachtová, Štěpánka (referee) ; Španová, Alena (advisor)
Polymerase chain reaction (PCR) is a molecular method which allows in vitro replication of nucleic acids. It allows the identification and quantification of microorganisms or to prove specific gene sequentions in different matrices of biological origin. Some nonpathogenic species of genus Clostridium cause damages of cheeses, so their identification and quantification is very important in cheesemaking. In this thesis, specific primers for genus Clostridium were tested. Bacterial DNA from culture collection strains and from strains isolated from damaged cheeses were used. Genus-specific region for Clostridium was amplified using specific primers. The PCR products (619 bp) were detected using electrophoresis in 1,8% agarose gel. Genus-specific character of primers was confirmed. DNA of Lactobacillus was used for negative control.
Identification of probiotic lactobacili in dairy products
Dofková, Květoslava ; Rittich, Bohuslav (referee) ; Španová, Alena (advisor)
The genus Lactobacillus is part of intestinal microbiota which constitution plays important role in health and well-being of the host. Probiotic properties of lactobacili have been studied in lot of studies and clinical effects (immune enhancement, prevention of diarrhoeal disease etc.) of some probiotic species have been documented. Therefore bacteria of genus Lactobacillus are used in the production of fermented dairy products and probiotic preparations. PCR method based on amplification of 16S – 23S rRNA intergenic spacer regions was developed for identification of Lactobacillus species presented in dairy products. Whole DNA obtained by phenol extraction from dairy product Actimel Natur was used in this work as template for PCR. Purified DNA was amplificated by genus-specific PCR. Bacteria of genus Lactobacilllus were detected due to the detection of specific PCR product. Using species-specific PCR probiotic species Lactobacillus casei was detected in the Actimel Natur in quantity that approximately corresponded to declared amounts.
Identification of selected probiotic bacteria in food additives
Bubeníková, Lucia ; Španová, Alena (referee) ; Rittich, Bohuslav (advisor)
Bifidobacteria and lactobacilli are gram-positive bacteria which belong to the lactic acid bacteria group. They are ordinary constituents of gastrointestinal microflora, play a pivotal role in human nutrition and health, prevent pathogen colonization and maintain normal mucosal immunity. In present days they are used in production of functional food products and pharmaceutical additives. Polymerase chain reaction (PCR) is used to the detection and identification of bacteria which belong to the genus of Bifidobacterium and Lactobacillus. Specific primers for each genus are used in PCR reactions for in vitro amplification of definite part of DNA. In this work, total DNA was isolated from three additives (Pangamin Bifi plus, Biopron Junior and Probiodom) by the method of phenol extraction and amplified using genus-specific PCR primers giving amplicons of 523 bp (Bifidobacterium) and 250 bp (Lactobacillus). Occurrence of bacteria of both genera was proved in all of the three tested probiotic products.
Identification of lactic acid bacteria in food products using amplification methods
Jurečková, Nela ; Trachtová, Štěpánka (referee) ; Španová, Alena (advisor)
The aim of the study was isolation of genomic DNA from 7 milk products (yogurts/ yogurt milk) in quality for use it in PCR. Magnetic microparticles (P(HEMA-co¬-GMA)) were used for the DNA isolation. DNA was reversibly adsorbed to the microspheres in the presence of 8% poly(ethylene glycol) PEG 6000 and 5 M sodium chloride. The adsorbed DNA was released from the microspheres in a low ionic strength TE buffer and use as DNA matrix in genus specific PCR Lactobacillus. The presence of DNA of genus Lactobacillus was detected in all analysed products.
Identification of lactic acid bacteria in probiotic products (tablets) using amplification methods
Balogová, Petra ; Trachtová, Štěpánka (referee) ; Španová, Alena (advisor)
Lactobacillus have been playing a crucial role in the fermentation processes for millenia. Their probiotic effects have been studied deeply and nowadays they are used in the production of fermented products and food adjunkt including probiotic preparations. This thesis was focused on bakteria of genus Lactobacillus in probiotic preparations (tablets). Polymerase chain reaction method (PCR) based on amplification of DNA was used to identify lactobacillus in seven probiotic preparations. As DNA matrices whole DNA recovered from tablets with the help of magnetic microparticles (P(HEMA-co¬-GMA) coated by carboxylic groups was used into a PCR mixture. Genus specific PCR products (250 bp) were detected with the help of gel electrophoresis on agaróse. DNA of genus Lactobacillus was proved in all but one product. In this product only probiotic bacteria were declared presence without family identification was proved. It contained different bakteria then bakteria of Genus Lactobacillus probably.

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