Course detail

Laboratory Classes in Molecular Biotechnology

FCH-MAO_MOB_PAcad. year: 2012/2013

Safety of work in microbiological and biotechnological laboratory. Arrangement of PCR laboratory. Cultivation of Lactobacillus gasseri cells for isolation of chromosomal DNA. Cell lysis. Removal of proteins using phenol extraction. Precipitation of DNA with ethanol. Verification of DNA integrity using agarose gel electrophoresis. Spectrophotometric estimation of concentration and purity verification of DNA. Preparation of PCR mixture for amplification of Lactobacillus gasseri using purified DNA as DNA matrix. Negative controls. PCR amplification. Detection of PCR products using agarose gel electrophoresis. Estimation of length of amplified fragment using external DNA standards. Sensitivity of PCR. Isolation of DNA from dairy product (yoghurt) using magnetic particles. Identification of Lactobacillus DNA in dairy product (yoghurt) by real time PCR. Testing of production of antimicrobial agents by probiotic strain Lactobacillus gasseri using cultivation methods and molecular diagnostic ones. Test.

Language of instruction

English

Number of ECTS credits

3

Mode of study

Not applicable.

Guarantor

doc. RNDr. Alena Španová, CSc.

Department

Institute of Food Science and Biotechnology (ÚCHPBT)

Offered to foreign students

Of all faculties

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

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

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 microbiological and biotechnological laboratory. Arrangement of PCR laboratory. 2. Cultivation of Lactobacillus acidophilus cells for isolation of chromosomal DNA. Cell lysis. 3. Removing of proteins using phenol extraction. Precipitation of DNA with ethanol. 4. Determination of DNA integrity using agarose gel electrophoresis. 5. Spectrophotometric estimation of concentration and purity of DNA. 6. Preparation of PCR mixture for genus specific amplification of Lactobacillus acidophilus using purified DNA as DNA matrix. Negative controls. 7. PCR amplification. Detection of PCR product using agarose gel electrophoresis. 8. Estimation of length of amplified fragment using external DNA standards. 9. Sensitivity of PCR. 10. Isolation of DNA from dairy product (yoghurt) . 11. Identification of Lactobacillus DNA in dairy product (yoghurt ) by PCR. Gel electophoresis of PCR amplicons. 12. -13. Test - identifikation of Lactobacillus cells in unknown sample using PCR.


Work placements

Not applicable.

Aims

The laboratory course cover methodologies used in DNA analysis

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

Sambrook F., Russell R. W.: Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York 2001. (EN)

Recommended reading

Not applicable.

Classification of course in study plans

  • Programme NPCP_CHM_INT Master's

    branch NPCO_CHM , 1 year of study, winter semester, elective

  • Programme CKCP_CZV lifelong learning

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

Type of course unit

 

Laboratory exercise

26 hod., compulsory

Teacher / Lecturer

doc. RNDr. Alena Španová, CSc.