SubjectsSubjects(version: 901)
Course, academic year 2022/2023
  
Methods for gene expression determination - MB140C76
Title: Metody stanovení genové exprese
Czech title: Metody stanovení genové exprese
Guaranteed by: Department of Genetics and Microbiology (31-140)
Faculty: Faculty of Science
Actual: from 2022
Semester: winter
E-Credits: 3
Examination process: winter s.:
Hours per week, examination: winter s.:0/4 C [days/semester]
Capacity: 18
Min. number of students: 10
Virtual mobility / capacity: no
State of the course: taught
Language: Czech
Level: specialized
Note: enabled for web enrollment
Guarantor: RNDr. Tomáš Mašek, Ph.D.
Teacher(s): RNDr. Tomáš Mašek, Ph.D.
Mgr. Kristina Roučová
Mgr. Václav Vopálenský, Ph.D.
Class: Ultracentrifuga a 3 rotory
Gradientový PCR cycler
Opinion survey results   Examination dates   Schedule   
Annotation -
Last update: RNDr. Tomáš Mašek, Ph.D. (22.08.2017)
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.
Literature -
Last update: RNDr. Tomáš Mašek, Ph.D. (22.08.2017)

No original textbooks are available. All manuals, protocols and other directions will be provided during the course.

Requirements to the exam -
Last update: RNDr. Tomáš Mašek, Ph.D. (22.08.2017)

Students must pass examination test and provide correct working protocols. Other information including recommended literature, presentations and protocols are available on the webpage (http://web.natur.cuni.cz/~pospisek).

Syllabus -
Last update: RNDr. Tomáš Mašek, Ph.D. (22.08.2017)

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.

 
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