Bi4030cen Laboratory Course in Molecular Biology

Faculty of Science
Spring 2021

The course is not taught in Spring 2021

Extent and Intensity
0/3/0. 3 credit(s). Type of Completion: z (credit).
Teacher(s)
Mgr. Tibor Botka, Ph.D. (seminar tutor)
Guaranteed by
Mgr. Tibor Botka, Ph.D.
Department of Experimental Biology - Biology Section - Faculty of Science
Contact Person: Mgr. Tibor Botka, Ph.D.
Supplier department: Department of Experimental Biology - Biology Section - Faculty of Science
Prerequisites
(! Bi4030c Molecular biology - pract. ) && ! NOWANY ( Bi4030c Molecular biology - pract. )
Basic knowledge of general genetics, microbiology, inorganic chemistry, organic chemistry and biochemistry.
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
Course objectives
This practical course is conceived as an introduction to methods of molecular biology. At the end of the course, students should be able to analyze genetic material including chromosomal DNA and perform cloning experiments with bacterial plasmids.
Learning outcomes
Student will be able to:
isolate plasmid DNA of sufficient quality and quantity;
analyze isolated genetic material by basic methods of molecular biology such as restriction endonuclease cleavage and ELFO;
prepare sufficient amount of PCR amplicon of the gene of interest;
use the isolated plasmid and the prepared PCR product to create a recombinant vector;
transfer the construct to E. coli cells and select transformants;
use gained knowledge in subsequent laboratory practice.
Syllabus
  • 1. Preparation of solutions and stock enzymes used in molecular biology.
  • 2. Isolation of plasmid DNA (pUC series, pBluescript). Determination of DNA concentration and purity.
  • 3. Polymerase chain reaction (PCR)
  • 4. Restriction analysis of nucleic acids, agarose gel electrophoresis.
  • 5. Restriction mapping, isolation of DNA from agarose gels.
  • 6. DNA cloning in basic types of vectors, DNA transfer to prokaryotic cells, transformation, selection of clones.
  • 7. Centrifugation techniques, preparation of saccharose density gradients.
Literature
    required literature
  • Protocols in Study Materials in IS and PowerPoint presentations
    recommended literature
  • S. Carson, H. B. Miller, M. C. Srougi, D. S. Witherow. Molecular Biology Techniques (4th Ed.) - A Classroom Laboratory Manual. Academic Press, 2019. eBook ISBN: 9780128157756, Paperback ISBN: 9780128180242
Teaching methods
The practical course is taught in seven 5-hour blocks in a molecular biology laboratory. Students perform the methods according to pre-examined protocols.
Assessment methods
The prerequisite for successful completion of the course is participation in all practical exercises, elaboration of protocols from performed experiments and passing a written exam focused on practical questions.
Language of instruction
English
Further Comments
The course is taught annually.
The course is taught: every week.

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