BTLW100021 Exercises in molecular biology I

Type
course with continuous assessment
Semester hours
3
Lecturer (assistant)
Groß, Angelina Sarah , Wiltschi, Birgit , Klausberger, Miriam , Schoberer, Jennifer , Strasser, Richard , Grabherr, Reingard , Luschnig, Christian
Organisation
Biotechnology and Food Science
Offered in
Sommersemester 2026
Languages of instruction
Englisch
Content

The teaching material contains two examples that are intended to familiarise students with basic molecular biology methods.

In the ‘Epigenetics’ example, genomic DNA is purified from transgenic plants. The presence of specific DNA methylation patterns in the promoter of the transgene is detected using methylation-sensitive restriction enzymes and subsequent PCR (polymerase chain reaction). The copy number of the transgene is determined by quantitative PCR and compared with the amount of protein formed. This example focuses on several important methods: (i) standard PCR, which enables the specific amplification of certain DNA segments; (ii) quantitative PCR, which allows the relative or absolute quantification of certain DNA segments; (iii) the detection of proteins using Western blot; and (iv) DNA sequence analysis using various databases.

In the ‘E. coli’ experiment, students learn how to use a range of enzymes that are used to modify DNA segments, making them easier to work with in the laboratory. Students also learn how to use the model organism E. coli and its application in molecular biology work. This example focuses on the following important methods: (i) use of restriction endonucleases to cut DNA, (ii) manipulation of linearised DNA fragments, (iii) creation of so-called competent cells, (iv) transformation of E. coli, (v) blue-white screening using the lacZ gene, (vi) analysis of transformants using agarose gel electrophoresis.

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Previous knowledge expected

BTLW100013 Allgemeine Mikrobiologie Übungen
Objective (expected results of study and acquired competences)

By the end of the training the students will

- understand the basic methods of cloning a reporter gene into a bacterial plasmid
- can perform in silico cloning
- can interpret the results of ligation and E. coli transformation
- can calculate transformation efficiency
- can examine the success of cloning using PCR, plasmid purification, restriction digestion and agarose gel electrophoresis
- can determine the expression of a reporter gene by measuring fluorescence emission
- recognise the critical factors that influence the expression of a gene in bacteria
- can evaluate the results obtained from cloning and gene expression by comparative reference
- perform quantitative PCR to quantify DNA fragments
- have a basic knowledge of methods for purifying genomic DNA from eukaryotic cells
- have a basic understanding of epigenetic processes in eukaryotic cells
- are familiar with the theory and practical implementation of polymerase chain reaction (PCR)
- can use the Western blot method to detect specific proteins
- can perform comparative DNA sequence analyses using databases
- can question the results and their practical relevance
You can find more details like the schedule or information about exams on the course-page in BOKUonline.