Analýza DNA Lactobacillus s využitím PCR v reálném čase a HRM analýzy
| Autor: | Aksamitová, Dagmar |
|---|---|
| Jazyk: | čeština |
| Rok vydání: | 2016 |
| Předmět: |
polymerázová řetězová reakce (PCR)
polymerase chain reaction (PCR) fluorescent dye rod Lactobacillus identifikace laktobacilů DNA isolation high resolution melt analysis (HRMA) izolace DNA fluorescenční barvivo vysokorozlišovací analýza křivek tání (HRMA) genus Lactobacillus lactobacilli identification |
| Druh dokumentu: | masterThesis |
| Popis: | The rapid and accurate identification of the bacterium of the genus Lactobacillus, which are an important part of the normal gastrointestinal microflora and fermented dairy products are currently mainly used amplification methods. The aim of the study was to analyze the possibility of resolution of selected bacterial strains of the genus Lactobacillus, using the metod of polymerase chain reaction in the real time combined with high resolution melting curve analysis (qPCR HRM). It was tested five primers designed for qPCR-HRM analysis of lactic acid bacteria. The specificity of the primers was also verified simultaneously using bioinformatic analysis. On the basis of analysis of the DNA were selected as the most appropriate primers P1V1/P2V1, V3F/V3R and V6F/V6R. The suitability of the primers V3F/V3R and V6F/V6R was verified on a complex sample of food supplement from which the DNA was isolated using magnetic particles. The presence of bacteria of the genus Lactobacillus was performed using high resoluting melting analysis curves. The obtained results were in agreement with the information given by the manufacturer. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
| Externí odkaz: |
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