Bi7430c Molecular biotechnology - practical course

Faculty of Science
Autumn 2004
Extent and Intensity
0/4/0. 4 credit(s). Type of Completion: z (credit).
Teacher(s)
doc. Ing. Bohuslav Rittich, CSc. (seminar tutor)
doc. RNDr. Alena Španová, CSc. (seminar tutor)
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
Prerequisites (in Czech)
NOW( Bi7430 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 and molecular biotechnical laboratory. GMO. Cultivation of bacterial E. coli JM109(pUC19)cells. Microisolation of plasmid DNA pUC19.Cultivation of bacterial E. coli JM109(pUC19::dim1)cells. Isolation of recombinant DNA pUC19::dim1. Linearisation of plasmid DNA with restrictase EcoRI. Gel electrophoresis of DNA and estimation of restriction fragment lenght. Transfer of recombinant and nonrecombinant plasmid DNAs into bacterial cells E. coli JM109 using electrotransformation. Plating and selection of electrotransformants. Evaluation of electrotransformation. Preparation of DNA probe from plasmid DNA - neradioactive labelling with digoxigenine. Dot blot. Southern blot. DNA/DNA hybridization at high stringent conditions. Immunological detection of hybridization products.
Syllabus
  • 1. Safety of work in and molecular biotechnical laboratory. GMO. 2. Cultivation of bacterial E. coli JM109(pUC19)cells. 3. Microisolation of plasmid DNA pUC19. 4. Cultivation of bacterial E. coli JM109(pUC19::dim1)cells. 5. Isolation of recombinant DNA pUC19::dim1. 6. Linearisation of plasmid DNA with restrictase EcoRI. Gel electrophoresis of DNA and estimation of restriction fragment lenght. 7. Transfer of recombinant and nonrecombinant plasmid DNAs into bacterial cells E. coli JM109 using electrotransformation. 8. Plating and selection of electrotransformants carrying recombinant and nonrecombinant plasmid DNA. Evaluation of electrotransformation. 9. Preparation of DNA probe from plasmid DNA - neradioactive labelling with digoxigenine. 10. Dot blot. 11. Southern blot. 12. DNA/DNA hybridization at high stringent conditions. 13. Immunological detection of hybridization products. 14. Evaluation of hybridization.Test. 15. Evaluation of protocols and test.
Literature
  • G. C. Saunders, H. C. Parkers. Analytical Molecular Biology. RSC. Cambridge 1999.
  • F. Sambrook, R.W. Russell Molecular Cloning. A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press. 2001.
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 2005, Autumn 2006, Autumn 2007, Autumn 2008, 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 2004, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2004/Bi7430c