Bi7430c Molecular biotechnology - practical course

Faculty of Science
Autumn 2008
Extent and Intensity
0/4/0. 4 credit(s). Type of Completion: z (credit).
Teacher(s)
doc. RNDr. Alena Španová, CSc. (seminar tutor)
doc. Ing. Bohuslav Rittich, CSc. (seminar tutor)
RNDr. Marie Vojtíšková, CSc. (seminar tutor), doc. RNDr. Alena Španová, CSc. (deputy)
Guaranteed by
doc. RNDr. Alena Španová, CSc.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: doc. RNDr. Alena Španová, CSc.
Timetable
Thu 8:00–11:50 Bpb,02012
  • Timetable of Seminar Groups:
Bi7430c/01: No timetable has been entered into IS. B. Rittich, A. Španová, M. Vojtíšková
Prerequisites
NOW( Bi7430 Molecular biotechnology )
The course is for the students with practical interest in selected laboratory methods 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 12 student(s).
Current registration and enrolment status: enrolled: 0/12, only registered: 0/12, only registered with preference (fields directly associated with the programme): 0/12
fields of study / plans the course is directly associated with
Course objectives
Safety of work in molecular biotechnical laboratory. Word with GMO. Cultivation of bacterial cells E. coli carrying recombinant and nonrecombinant plasmid pUC19 DNA. Microisolation of plasmid DNA and recombinant plasmid DNA estimation of their lenghts and the lenght of insert.Transfer of reombinant and nonrecombinant plasmid DNA into bacterial cells E. coli JM109. Plating and selection of transformants. Preparation of DNA probe labelled with digoxigenine, performance of DNA/DNA hybridization (dot blot and Southern blot) and immunological detection of hybridization products. Evaluation of results. The practical course is supplemented by course Bi7430.
At the end of this course, each student should be able to known some procedures of molecular biotechnology. There are included methods of microisolation of plasmid DNA, estimation of insert length,, transformation of plastid DNA into bacterial cells and selection of transformants. In the next part of the course students will prepare DNA probe and performe DNA/DNA hybridisation including imunological detection of hybridisation product.
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.
Assessment methods
Training is enclosed by a test.
Language of instruction
Czech
Further comments (probably available only in Czech)
The course is taught annually.
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 2009, Autumn 2010, Autumn 2011, Autumn 2011 - acreditation, Autumn 2012, Autumn 2013, Autumn 2014, Autumn 2015, Autumn 2016, autumn 2017, Autumn 2018, Autumn 2019, Autumn 2020, autumn 2021, Autumn 2022, Autumn 2023, Autumn 2024.
  • Enrolment Statistics (Autumn 2008, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2008/Bi7430c