Bi7430c Molecular biotechnology - practical course

Faculty of Science
Autumn 2012
Extent and Intensity
0/4/0. 4 credit(s). Type of Completion: z (credit).
Teacher(s)
prof. RNDr. Zbyněk Prokop, Ph.D. (seminar tutor)
Mgr. Šárka Nevolová, Ph.D. (seminar tutor)
Mgr. Pavel Dvořák, Ph.D. (seminar tutor)
Mgr. Lukáš Chrást (seminar tutor)
Guaranteed by
prof. Mgr. Jiří Damborský, Dr.
Department of Experimental Biology - Biology Section - Faculty of Science
Contact Person: prof. RNDr. Zbyněk Prokop, Ph.D.
Supplier department: Department of Experimental Biology - Biology Section - Faculty of Science
Prerequisites
NOW ( Bi7430 Molecular biotechnology )
The course is destined for the students with practical interest in selected laboratory procedures of molecular biotechnology.
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 24 student(s).
Current registration and enrolment status: enrolled: 0/24, only registered: 0/24, only registered with preference (fields directly associated with the programme): 0/24
fields of study / plans the course is directly associated with
Course objectives
At the end of the course students should be able to explain principles and utilize selected procedures of molecular biotechnology, e.g. methods of E. coli cultivation, microisolation of plasmid DNA, methods of bacterial cell transformation and selection of transformants carrying both non-recombinant and recombinant plasmid DNA, DNA cleavage using restrictases, and estimation of restriction fragments. During the course the students will learn the Southern blot, dot blot, DNA probe preparation, and DNA/DNA hybridisation including immunological detection of hybridisation products.
Syllabus
  • 1. Safety of work in an molecular biotechnology laboratory. Work withGMO.
  • 2. Cultivation of bacterial E. coli JM109, E. coli JM 109 (pUC19) a E. coli JM109 (pUC19::dim1) .
  • 3. Microisolation of plasmid DNA pUC19 and recombinant plasmid DNA pUC19::dim1.
  • 4. Estimation of plasmid DNA isolation using agarose gel electrophoresis.
  • 5. Transfer of recombinant and nonrecombinant plasmid DNA into competent bacterial cells E. coli JM109
  • 6. Plating and selection of transformants carrying recombinant and nonrecombinant plasmid DNA.
  • 7. Evaluation of transformation.
  • 8.Cleavage of plasmid DNAs with restrictase EcoRI (linearisation) and BamHI.
  • 9. Gel electrophoresis of DNA and estimation of restriction fragment lenghts using lambda/HindIII standards.
  • 10. Southern blot. Preparation of DNA for dot blot hybridisation.
  • 11. Preparation of DNA probe from plasmid DNA - neradioactive labelling with digoxigenine.
  • 12. DNA/DNA hybridization at high stringent conditions.
  • 13. Immunological detection of hybridization products.
  • 14. Evaluation of results of hybridization.Test.
Literature
  • F. Sambrook, R.W. Russell Molecular Cloning. A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press. 2001.
  • G. C. Saunders, H. C. Parkers. Analytical Molecular Biology. RSC. Cambridge 1999.
Teaching methods
During the laboratory course, each student works independently and processes his own sample. They must evaluate the results of their experiments in protocols. The protocols (4 in number) are elaborated in the form of a poster according to the following scheme: Introduction, Aim of work, Material and methods, Results, Discussion, Conclusion. In the protocol each student describes, compares, analyses, evaluates, and discusses his or her own results.
Assessment methods
The course is closed by a course-unit credit. The students obtain their credits for active attendance at the course and for the elaboration of protocols (4).
Language of instruction
Czech
Further comments (probably available only in Czech)
Study Materials
Course is no more offered.
The course is taught: every week.
Information on course enrolment limitations: Přednost mají studenti specializace mikrobiologie.
The course is also listed under the following terms Autumn 2007 - for the purpose of the accreditation, Autumn 2010 - only for the accreditation, Autumn 2003, Autumn 2004, Autumn 2005, Autumn 2006, Autumn 2007, Autumn 2008, Autumn 2009, Autumn 2010, Autumn 2011, Autumn 2011 - acreditation, Autumn 2013, Autumn 2014, Autumn 2015, Autumn 2016, autumn 2017, Autumn 2018, Autumn 2019, Autumn 2020.
  • Enrolment Statistics (Autumn 2012, recent)
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