Course detail

Laboratory Classes in Molecular Biotechnology

FCH-MC_MOB_PAcad. year: 2015/2016

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

Czech

Number of ECTS credits

2

Mode of study

Not applicable.

Guarantor

doc. RNDr. Alena Španová, CSc.

Department

Institute of Food Science and Biotechnology (ÚCHPBT)

Learning outcomes of the course unit

Practical aspects of various experimental techniques used in DNA research

Prerequisites

It is necessary pass out lectures Bioanalytical methods.

Co-requisites

Not applicable.

Planned learning activities and teaching methods

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.

Assesment methods and criteria linked to learning outcomes

Laboratory course is finished by identification of LAB using PCR

Course curriculum

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.

Work placements

Not applicable.

Aims

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.

Recommended optional programme components

Not applicable.

Prerequisites and corequisites

Not applicable.

Basic literature

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

Recommended reading

Not applicable.

Classification of course in study plans

  • Programme NPCP_CHTP Master's

    branch NPCO_CHTP , 1. year of study, winter semester, compulsory-optional

  • Programme NKCP_CHTP Master's

    branch NKCO_CHTP , 1. year of study, winter semester, compulsory-optional

  • Programme CKCP_CZV lifelong learning

    branch CKCO_CZV , 1. year of study, winter semester, compulsory-optional

Type of course unit

 

Laboratory exercise

26 hours, compulsory

Teacher / Lecturer

RNDr. Aleš Kovařík, CSc.
doc. RNDr. Alena Španová, CSc.
Ing. Štěpánka Trachtová, Ph.D.