|
Lecturer(s)
|
-
Šafář Jan, RNDr. Ph.D.
-
Šimková Hana, Ing. CSc.
-
Šafářová Dana, Mgr. Ph.D.
|
|
Course content
|
Preparation of high-molecular-weight DNA. Construction of a BAC DNA library. Characterization of the DNA library - number of clones, efficiency of transformation, insert size analysis. Use of an automatic workstation for handling with the libraries - picking, gridding, preparation of library pools. Screening of a BAC library by PCR on pokos. Quantitative PCR - principle, detection of GMO in selected samples. Cloning of PCR fragment - preparation of DNA fragment, ligation into pGem-T plasmid, transformation, screening of colonies.
|
|
Learning activities and teaching methods
|
|
Dialogic Lecture (Discussion, Dialog, Brainstorming), Observation, Demonstration, Laboratory Work
|
|
Learning outcomes
|
DNA cloning, construction of BAC DNA library and its screening. Quantitative PCR.
Student is able to (after attending the course): - Prepare the plasmid DNA. - Prepare the clones. - Work with DNA libraries and DNBA clones. - Browse the genomic DNA libraries using DNA.
|
|
Prerequisites
|
unspecified
|
|
Assessment methods and criteria
|
Student performance
Active attendance, protocols elaboration.
|
|
Recommended literature
|
-
Brdička, R. Lidský genom na rozhraní tisíciletí. Grada Publishing, Praha, 2001.
-
Cantor, C.R., Smith, C.L. Genomics: The Science and Technology Behind the Human Genome Project, John Wiley and Sons, New York, 1999..
-
Cullis, C.A. Plant Genomics and Proteomics. John Wiley and Sons, Inc., New York, 2004.
-
Primrose, S.B. Principles of Genome Analysis: A guide to mapping and sequencing DNA from different organisms. Blackwell Science Inc., 1998..
|