Course detail
Laboratory Classes in Molecular Biotechnology
FCH-MC_MOB_PAcad. year: 2025/2026
The course is focused on practical mastering of method polymerase chain reaction in identification of bacteria of genus Lactobacillus in milk product (probiotic drink). PCR-ready DNA is isolated from the product using magnetic microspheres. PCR mixtures are prepared after estimation of DNA concentration and quality. DNA is amplified in conventional PCR and in real-time PCR. Working out of for protocols in the form of posters is part of the course.
Language of instruction
Number of ECTS credits
Mode of study
Guarantor
Entry knowledge
Rules for evaluation and completion of the course
Attendance in laboratory course is obligatory.
Aims
Practical knowledges of some basis experimental techniques used in DNA research.
Study aids
Prerequisites and corequisites
Basic literature
Recommended reading
Classification of course in study plans
- Programme NPCP_CHMA Master's
specialization CHBL , 1 year of study, winter semester, compulsory-optional
- Programme NPCP_PCHBT Master's 1 year of study, winter semester, compulsory-optional
Type of course unit
Guided consultation in combined form of studies
Teacher / Lecturer
Laboratory exercise
Teacher / Lecturer
Syllabus
2. Cultivation of Lactobacillus cells for isolation of bacterial DNA. Estimation of purity of bacterial culture.
Cell lysis and preparing of crude cell lysate. Removing of proteins using phenol extraction. Precipitation of DNA with ethanol. Determination of DNA integrity using agarose gel electrophoresis. Preparation of a protocol in the form of poster.
3. Spectrophotometric estimation of DNA concentration and purity. Preparation of PCR mixture for amplification of specific part of Lactobacillus DNA using purified DNA. Preparation of negative control. PCR amplification. Detection of PCR product using agarose gel electrophoresis. Estimation of length of amplicon with help of DNA standard and programme Bionumerics. Preparation of a protocol in the form of poster.
4. Isolation of whole DNA from crude cell lysates of dairy product (probiotic drink) using magnetic particles P(HEMA-co-GMA). Amplification of DNA by PCR in real time. Preparation of a protocol in the form of poster.
5. Introduction to bioinformatics - lecture a practical excersize on PC.
6. Test and control of protocols.