Personal Details

Der Kurs „Einführung in molekularbiologische Techniken“, führt Studierende in grundlegende molekularbiologische Techniken ein. Er behandelt Restriktionsenzymverdauung, Agarose-Gelelektrophorese und DNA-Konzentrationsbestimmung mittels Spektrophotometrie. Die Studierenden sammeln praktische Erfahrung bei der Durchführung dieser Techniken, der Analyse experimenteller Daten und der Erstellung einer Restriktionskarte eines unbekannten Plasmids. Der Kurs legt den Schwerpunkt auf sichere Laborpraktiken, einschließlich des Umgangs mit Enzymen, der Abfallentsorgung und der richtigen Pipettiertechniken mit automatischen Pipetten. Desweiteren erfolgt die Erfassung der Ergebnisse in Form eines wissenschaftlichen Protokolls („lap report“) in englischer Sprache.

Aktivierende Methoden

Endprüfung

Protokoll in englischer Sprache, Mitarbeit

- Köhlern des Mikroskops und Kalibrierung des Messokulars

- Durchführung einer Plasmolyse (zB Zwiebel) und mikroskopische Auswertung

- Färbung und Analyse zum Stärkenachweis (zB Kartoffel, Banane, Weizen)

- Färbung und Analyse von Zellen der Mundschleimhaut

- Mikroskopische Analyse von Teichwasser

- Präparation und Mikroskopie von Chromoplasten in Pflanzengewebe (zB Karotte)

- Anfertigen und Mikroskopie der pflanzlichen Zellwand (verholzt, nicht verholzt; mit Färbung)

- Anfertigen und Mikroskopie eines Blattquerschnitts

- Mikroskopie eines menschlichen Haares, sowie fertiger Präparate (Blutausstrich, Mitosestadien einer Zwiebel, Hautpräparate, etc)

Dokumentation aller mikroskopischer Analysen und Messen mittels Messokular bzw. Kamera.

Praktikum, Präsenzlehre (Vorbesprechung), Fernlehre

Immanente Leistungsüberprüfung

Die Gesamtnote ergibt sich aus dem Engagement während des Praktikums, der Beurteilung der Qualität der im Praktikum anzufertigenden Protokolle, sowie der Abgabe eines Arbeitsauftrages.

- Introduction to Genetic Analysis Methods:

- Explanation and practical implementation of various genetic analysis techniques.

- Reverse Transcriptase Polymerase Chain Reaction (RT-PCR):

- Detection of leukemia-associated fusion transcripts.

- Gene Amplification and Hybridization Techniques:

- Use of immobilized, allele-specific oligonucleotides for mutation detection in the cystic fibrosis gene.

- Real-Time Polymerase Chain Reaction (RT-PCR):

- Identification of BRAF and KRAS mutations in cancer cells.

- Categorization of patients with poorer prognosis based on these mutations.

- Breast Cancer Clinical Classification:

- Classification based on the expression of:

- Estrogen and progesterone receptors (ER and PR).

- Human epidermal growth factor receptor 2 (HER2).

- Percentage of Ki67-positive cancer nuclei.

- Immunohistochemistry:

- Analysis of selected factors.

- Examination of expression patterns in various molecular subtypes of breast cancer.

- The theoretical basis of each of the analyses conducted in the laboratory is explained in a preceding seminar.

- Students conduct genetic analyses according to detailed reports provided by the lecturers.

- Laboratory results are discussed with lecturers at the end of each unit and are summarized in a report whose structure corresponds to that of a scientific paper.

Immanente Leistungsüberprüfung

presence, motivation, participation, practical skills (results), written report.

- Deepening of theoretical immunological knowledge and practical application to research problems from the field of allergy research

- Working in small teams of 2 to 3 people on a scientific research question

- Finding as a team the best way to answer the research question

- Designing and performing of experiments using state-of-the-art immunological and molecular biological methods such as ELISAs, SDS-PAGE, immunoblotting, PCR, microscopy and flow cytometry

- Writing of laboratory report in the format of a scientific publication, in which the background of the research topic is summarized and the experimental work, the results are described and discussed (each student writes a lab report)

- Dialogue and discussion

- Individual work

- Group work

- Supervised distance learning

- Self-study

- Problem-based learning

- Project work

- Research-based learning

Immanente Leistungsüberprüfung

Assessment of the design of the experiments; assessment of the performance of the experiments; assessment of the quality of the laboratory report

- General RNA Handling Procedures

- Experiment 1: Northern Blot: This experiment investigates the differential expression of the GAL1 gene in yeast (Saccharomyces cerevisiae) grown in media containing either glucose or galactose. (glucose/galactose metabolism in yeast, RNA extraction from yeast, denaturing RNA agarose gel, RNA transfer, specific oligonucleotide hybridization, band detection, quantitative PCR)

- Experiment 2: Band Shift – EMSA: This experiment focuses on detecting the interaction between human Y RNA and the La protein using the Electrophoretic Mobility Shift Assay (EMSA), also known as a band shift assay. (in vitro transcription with T7 RNA polymerase, RNA purification, RNA folding, native poly-acrylamide gel electrophoresis, RNA staining using methylene blue, detection of RNP complexes)

- Experiment 3: RNA Stability: This experiment explores the inherent instability of RNA compared to DNA. (temperature dependence, pH dependence, RNAses)

Laboratory

Immanente Leistungsüberprüfung

presence, motivation, participation, practical skills (results), written report.

- Objective: Estimate the toxicological and therapeutic potential of a small molecule drug using cell-based test systems.

- Focus Areas:

- Analyse the small molecule's activation potential on the heat shock response pathway.

- Assess possible cytotoxic effects in a concentration-dependent manner.

- Assays and Methods Available:

- Luciferase reporter assays

- Western blot

- qPCR

- Flow cytometry

- ELISA

- General viability assays

- Student Role: Select and apply suitable methods from the provided options.

Practical course with independent performance of the experiments

Immanente Leistungsüberprüfung

Entry exam on the computer (multiple choice), evaluation of performance in the lab, protocol