Czech

Not applicable.

Practical aspects of various experimental techniques used in DNA research

It is necessary pass out lectures Bioanalytical methods.

Not applicable.

The course uses teaching methods in form of Laboratory exercise - 2 teaching hours per week. The e-learning system (LMS Moodle) is available to teachers and students.

Laboratory course is finished by identification of LAB using PCR

1. Safety of work in laboratory. Arrangement of PCR laboratory. Pipetting of small volumes. 2. Cultivation of Lactobacillus casei cells for isolation of bacterial DNA. Estimation of bacterial purity.
3. Cell lysis and preparing 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.
6. Spectrophotometric estimation of concentration and purity.
7. Preparation of PCR mixture for amplification of specific part of Lactobacillus DNA using purified DNA as DNA matrix. Negative control.
8. PCR amplification.
9. Detection of PCR product using agarose gel electrophoresis.
10. Estimation of length of amplicon using DNA standards and programme Bionumerics.
11. Isolation of whole DNA from crude cell lysates of dairy product (probiotic drink) using method of magnetic separation with magnetic particles P(HEMA-co-GMA).
12. Amplification of DNA Lactobacillus in dairy product (probiotic drink ) by RT-PCR.
13. Test and control of protocols.

Not applicable.

The aim of the laboratory course is to cover methodologies used in the work with DNA particularly its amplification in PCR. The next aim covers the preparation of poster.

Attendance in laboratory course is obligatory.

Not applicable.

Not applicable.

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

Not applicable.