Laboratory Classes in Molecular Biotechnology
FCH-MC_MOB_PAcad. year: 2021/2022
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.
Learning outcomes of the course unit
Practical knowledges of some basis experimental techniques used in DNA research.
It is recomended to pass out the lecture Bioanalytical methods.
Recommended optional programme components
Recommended or required reading
Španová A., Rittich B.: Analýza vybraných druhů baktérií mléčného kvašení pomocí metod molekulární biologie (CS)
Sambrook F., Russell R. W.: Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York 2001. (CS)
Planned learning activities and teaching methods
The teaching is realised in 4 blocks. Each block runs 6 hours every 2 weeks. Part of the teaching is one introductery and one concluding hour.
The teaching requires 26 hours alltogether.
E-learning system LMS Moodle is available for teachers and students.
Assesment methods and criteria linked to learning outcomes
Laboratory course is finished by test and cotrol of protocols.
Language of instruction
1. Safety of the work in laboratory. Arrangement of PCR laboratory. Pipetting of small volumes.
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.
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.
Specification of controlled education, way of implementation and compensation for absences
Attendance in laboratory course is obligatory.
Classification of course in study plans
- Programme NKCP_CHTPO Master's
specialization BCH , 1. year of study, winter semester, 2 credits, compulsory-optional
- Programme NPCP_CHTP Master's
branch NPCO_CHTP , 1. year of study, winter semester, 2 credits, compulsory-optional
- Programme NKCP_CHTP Master's
branch NKCO_CHTP , 1. year of study, winter semester, 2 credits, compulsory-optional
- Programme NPCP_CHMA Master's
specialization CHBL , 1. year of study, winter semester, 2 credits, compulsory-optional
- Programme NPCP_MA Master's
branch NPCO_MA , 1. year of study, winter semester, 2 credits, compulsory-optional