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Preparation of PCR-ready DNA
Čuta, Robert ; Vojtíšková, Marie (referee) ; Rittich, Bohuslav (advisor)
In these days are probiotic lactic acid bacteria (LAB) very often used in food processing industry, such as milk products, cheese and fermented salami production. As well as the food conservation agent. Except of the industry usage of the LAB there are microbiological aspects. Identification of the bacterial species methods based on the isolation and amplification of DNA are very often used last few years. Diploma thesis deal with the bacterial cell of Lactobacillus species lysis and it´s optimalization. At first it was tested the optimal concentration of lysozyme (3mg/ml, 5mg/ml, 10mg/ml) and the exposure times (3, 5 a 10 hours). Another testing was aimed to the use and suitability of washing powder to the bacterial cell of Lactobacillus species lysis. I tested the Amway washing powder optimal concentration (1%, 2%, 3% a 4%). Four of another comercial washing powders were tested too. All these tests were performed at the pure Lactobacillus bacterial culture. To ensure the results I tested the washing powders at the real food matrix (Acidified milk, yogurt mango, yogurt white). All the methods were evaluated at the amplification method PCR with specific primers for the Lactobacillus genus. The DNA isolation was performed with the paramagnetic microsperes P(HEMA-co-GMA) and the amount of the DNA was quantified spectrophotometrically. The PCR products detection was performed with the agarose gel electrophoresis.
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Preparation of PCR-ready DNA
Čuta, Robert ; Vojtíšková, Marie (referee) ; Rittich, Bohuslav (advisor)
In these days are probiotic lactic acid bacteria (LAB) very often used in food processing industry, such as milk products, cheese and fermented salami production. As well as the food conservation agent. Except of the industry usage of the LAB there are microbiological aspects. Identification of the bacterial species methods based on the isolation and amplification of DNA are very often used last few years. Diploma thesis deal with the bacterial cell of Lactobacillus species lysis and it´s optimalization. At first it was tested the optimal concentration of lysozyme (3mg/ml, 5mg/ml, 10mg/ml) and the exposure times (3, 5 a 10 hours). Another testing was aimed to the use and suitability of washing powder to the bacterial cell of Lactobacillus species lysis. I tested the Amway washing powder optimal concentration (1%, 2%, 3% a 4%). Four of another comercial washing powders were tested too. All these tests were performed at the pure Lactobacillus bacterial culture. To ensure the results I tested the washing powders at the real food matrix (Acidified milk, yogurt mango, yogurt white). All the methods were evaluated at the amplification method PCR with specific primers for the Lactobacillus genus. The DNA isolation was performed with the paramagnetic microsperes P(HEMA-co-GMA) and the amount of the DNA was quantified spectrophotometrically. The PCR products detection was performed with the agarose gel electrophoresis.
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