Syllabus
Principles of the demonstrated methods, their advantages and limitations are explained in the theoretical part of the course. Experimental design, data evaluation and results interpretation will be also discussed.
In the practical part, we employ model gene reporter under the control of an inducible promoter.
Tasks: 1/ Principles of the work with RNA (preparation of solutions, disposables and other equipment). Total RNA isolation, concentration determination, RNA quality analysis using RNA denaturing agarose electrophoresis, DNaseI treatment, cDNA synthesis. 2/ RT PCR and qPCR approach comparison, absolute and relative quantification. 3/ Gene copy number determination by qPCR (data evaluation). 4/ Western blot 5/ Evaluation of inducible promoter activity by luciferase activity measurement and by cell growth determination.
Annotation
The course is composed of the theoretical and the practical parts. Pivotal part of the course is dedicated to qPCR and work with RNA. The most common experimental approaches for detection and quantification of gene expression in its different phases will be demonstrated on one gene model.
The course is intended for cell and molecular biology MSc. and PhD. students, but it is also suitable for those interested in gene expression determination methods from a wide variety of biological sciences. The course is organized as 3-day-turnus and is also accessible to english-speaking students.