PřF:B6721c Spec. Meth. Microorg. Anal. I.

B6721c Special Methods of Microorganisms Analysis I. - practice

Faculty of Science
Spring 2000

Extent and Intensity

0/4/0. 0 credit(s). Type of Completion: z (credit).

Teacher(s)

Guaranteed by

Prerequisites

The laboratory training course is a part of the course B6721.

Course Enrolment Limitations

The course is also offered to the students of the fields other than those the course is directly associated with.

fields of study / plans the course is directly associated with

Syllabus

The first part of laboratory training course is devoted to the basic methods of work with DNA, the second part is devoted to the polymerase chain reaction method (PCR) and its application to bacteria identification. Contents of laboratory training course is following: Cultivation of bacterial cells E. coli carrying plasmid pUC19. Microisolation of plasmid DNA. Purification of plasmid DNA - removal of RNA and proteins. Ethanol precipitation of DNA. Semiquantitative estimation of small amounts of plasmid DNA. Linearisation of plasmid DNA using restriction nuclease EcoRI. The estimation of linearised DNA length using programme Anagel. Cultivation of bacterial cells for the isolation of bacterial DNA. Isolation of bacterial DNA. Spectrophotometric estimation of purity and concentration of bacterial DNA. The preparation of PCR mixture for the specific amplification of Salmonella gene. Carry out of PCR on cycler. Detection of PCR product using agarose gel electrophoresis. PCR from the crude lysates of bacterial cells. PCR from one bacterial colony.

Literature

1. Sambrook J., Fritsch E.F., Maniatis T. (1989) Molecular Cloning. 2nd ed. CSHL Press. 2. Westermeier R. (1993) Electrophoresis in Practice. VCH Weinheim. 3. Dieffenbach C.W.and Dveksler G.S. (1995) PCR primer. CSHL Press.

Assessment methods (in Czech)

Cvičení je ukončeno zápočtem.

Language of instruction

Czech

Follow-Up Courses

Further Comments

The course is taught annually.
The course is taught: every week.