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Lecturer(s)
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Šafář Jan, RNDr. Ph.D.
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Šimková Hana, Ing. CSc.
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Hřibová Eva, Mgr. Ph.D.
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Šafářová Dana, Mgr. Ph.D.
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Course content
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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.
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Learning activities and teaching methods
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Dialogic Lecture (Discussion, Dialog, Brainstorming), Observation, Demonstration, Laboratory Work
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Learning outcomes
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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.
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Prerequisites
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unspecified
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Assessment methods and criteria
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Student performance
Active attendance, protocols elaboration.
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Recommended literature
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Brdička, R. Lidský genom na rozhraní tisíciletí. Grada Publishing, Praha, 2001.
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Cantor, C.R., Smith, C.L. Genomics: The Science and Technology Behind the Human Genome Project, John Wiley and Sons, New York, 1999..
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Cullis, C.A. Plant Genomics and Proteomics. John Wiley and Sons, Inc., New York, 2004.
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Primrose, S.B. Principles of Genome Analysis: A guide to mapping and sequencing DNA from different organisms. Blackwell Science Inc., 1998..
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