PřF:Bi6721c Spec.Meth.Microorg.Anal.I.-pr. - Course Information
Bi6721c Special Methods of Microorganisms Analysis I. - practical course
Faculty of ScienceSpring 2005
- Extent and Intensity
- 0/3/0. 3 credit(s). Type of Completion: z (credit).
- Teacher(s)
- doc. Ing. Bohuslav Rittich, CSc. (lecturer)
doc. RNDr. Alena Španová, CSc. (lecturer)
doc. Ing. Bohuslav Rittich, CSc. (seminar tutor)
doc. RNDr. Alena Španová, CSc. (seminar tutor) - Guaranteed by
- doc. Ing. Bohuslav Rittich, CSc.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: doc. Ing. Bohuslav Rittich, CSc. - Timetable of Seminar Groups
- Bi6721c/01: Thu 11:00–13:50 Bpb,02012, A. Španová
Bi6721c/02: Thu 8:00–10:50 Bpb,02012, A. Španová - Prerequisites (in Czech)
- NOW ( Bi6721 Spec. Meth. Microorg. Anal. I. )
- Course Enrolment Limitations
- The course is also offered to the students of the fields other than those the course is directly associated with.
The capacity limit for the course is 12 student(s).
Current registration and enrolment status: enrolled: 0/12, only registered: 0/12, only registered with preference (fields directly associated with the programme): 0/12 - fields of study / plans the course is directly associated with
- General Biology (programme PřF, B-BI, specialization Microbiology)
- General Biology (programme PřF, M-BI, specialization Microbiology)
- General Biology (programme PřF, N-BI, specialization Microbiology)
- Course objectives
- I. Princips of work with DNA on model of plasmid DNA. Cultivation of bacterial cells E.coli JM109(pUC19). Microisolation of plasmid DNA pUC19. Preparation of agarose gel, gel electrophoresis of DNA, visualisation of DNA on gel, documentation. Purification of DNA. Elimination of RNA using LiCl and RNase A, proteins elimination using phenol extraction, precipitation of DNA by ethanol. Linearisation of plasmid DNA with restrictase EcoRI and estimation of restriction fragment lenght using program Anagel. II. PCR Salmonella. Isolation and purification of bacterial DNA Salmonella typhimurium LB5000 and Escherichia coli JM109. Spectrophotometric estimation of DNA concentration and purity.Preparation of PCR mixture for specific amplification of gene of Salmonella. Carry out of PCR and detection of PCR product. PCR with crude cell lysate. Immunomagnetic separation (IMS)of cells Salmonella from milk. IMS-cultivation of Salmonella cells. IMS-PCR of Salmonella cells.
- Syllabus
- 1. Safety of work in microbiological and molecular biotechnological laboratory. Pipeting of small volumes of solvents using pippetmans. Preparation of media for cultivation of bacteria. 2. Cultivation of bacterial cells E.coli JM109 and E. coli JM109(pUC19). Estimation of the presence of plasmids using cultivation of cells on plates with antibiotic. 3. Microisolation of plasmid DNA. 4. Preparation of 1% agarose gel. Gel electrophoresis of DNA. Visualisation of DNA and gel documentation. 5. Purification of DNA. Removing of RNA using LiCl and RNase A.Removing of proteins using phenol extraction. 6. Precipitation of DNA with ethanol.Checking of RNA removing and estimation of small concentrations of DNA using gel electrophoresis with standards containing known amount of DNA. 7. Linearisation of plasmid DNA with restrction nuclease EcoRI. Gel electrophoresis of cleaved DNA with DNA standards of known length. 8. Estimation of length of restriction fragment of DNA. The use of program Anagel. 9. Preparation of Salmonella typhimurium LB5000 and Escherichia coli JM109 cells for isolation of chromosomal DNA. Deproteination of DNA using phenol extraction. Precipitation of DNA using ethanol. 10.Spectrophotometric estimation of concentration and purity of DNA. The estimation of DNA integrity using agarose gel electrophoresis. 11.Preparation of PCR mixture for specific amplification of gene Salmonella using purified DNA as DNA matrix. Negative control.PCR reaction. 12.Detection of PCR product using agarose gel electrophoresis. The characterisation of PCR laboratory. 13.Preparation of crude cell lysate for PCR.Preparation of PCR mixture using crude cell lysate as DNA matrix. Pozitive and negative controls. Provodení PCR a detekce PCR produktu. 14.Imunomagnetic separation (IMS)of cells Salmonella. IMS-cultivation (IMS-CM) of cells Salmonella. IMS-polymerase chain reaction (IMS-PCR)with cells Salmonella. 15.Identifikation of Salmonella cells in unknown sample using PCR.
- Literature
- F.Sambrook and D.W. Russell Molecular Cloning. A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press. 2001
- Assessment methods (in Czech)
- Cvičení je ukončeno zápočtem
- Language of instruction
- Czech
- Follow-Up Courses
- Further comments (probably available only in Czech)
- The course is taught annually.
Information on course enrolment limitations: Přednost mají posluchači specializace mikrobiologie.
- Enrolment Statistics (Spring 2005, recent)
- Permalink: https://is.muni.cz/course/sci/spring2005/Bi6721c