Czech

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Practical aspects of basic experimental techniques used in DNA analysis.Preparation of poster from own experimental results..

It is necessary pass out lectures Bioanalytical methods.

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Teaching methods depend on the type of course unit as specified in the article 7 of BUT Rules for Studies and Examinations.

There will be prepared 4 protocols based on own results in the form of a poster during the course. Laboratory course is finished by a test.

1. Safety of work in microbiological and biotechnological laboratory. Arrangement of PCR laboratory. 2. Cultivation of probiotic Lactobacillus gasseri culture for isolation of bacterial DNA. Estimation of purity of bacterial culture. 3. Cell lysis and preparation of crude cell lysate. 4. Removing of proteins using phenol extraction. Precipitation of DNA with ethanol. 5. Determination of DNA integrity using agarose gel electrophoresis. Visualisation of DNA and documentation of the gel.
6. Spectrophotometric estimation of concentration and purity of DNA.DNA dilution. 7. Preparation of PCR mixtures for specific amplification of DNA Lactobacillus using purified DNA as DNA matrix. Estimation of PCR sensitivity. Negative controls. 8. PCR amplification. Detection of PCR products using agarose gel electrophoresis. 9. Estimation of length of amplicons using external DNA standards and program Bionumerics.
10. Isolation of DNA from dairy product (probiotic drink) using magnetic separation with magnetic particles P(HEMA-co-GMA). 11. Identification and quantification of Lactobacillus DNA in dairy product (probiotic drink ) by PCR v reálném čase.
12. Testing of antimicrobial compounds production. Amplification of gene coding bacteriocin in probiotic strain L. gasseri.
13. Test.

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The laboratory course covers basic methodologies used in the work DNA and DNA amplification using polymerase chain reaction. The next aim is preparation of poster.

Attendance in laboratory course is obligatory.

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Španová A., Rittich B.: Analýza vybraných druhů baktérií mléčného kvašení pomocí metod molekulární biologie (CS)

Not applicable.