Bi9310 Real-time polymerase chain reaction

Faculty of Science
Autumn 2010
Extent and Intensity
2/0. 2 credit(s) (fasci plus compl plus > 4). Recommended Type of Completion: zk (examination). Other types of completion: k (colloquium).
Teacher(s)
doc. RNDr. Petr Vaňhara, Ph.D. (lecturer)
Mgr. Zdeněk Ručka, Ph.D. (assistant)
doc. RNDr. Sabina Ševčíková, Ph.D. (lecturer)
Guaranteed by
prof. RNDr. Jan Šmarda, CSc.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: doc. RNDr. Petr Vaňhara, Ph.D.
Prerequisites (in Czech)
Bi4035 Molecular biology - lab.pract. || Bi4020c Essential methods in mol. biol
Course Enrolment Limitations
The course is only offered to the students of the study fields the course is directly associated with.
fields of study / plans the course is directly associated with
Course objectives
At the end of this course, students should be able to:
understand PCR and real-time PCR principles
design a real-time PCR experiment, including template manipulation, chemistry and instrumentation
evaluate absolute and relative quantification data, SNP analysis and alelic discrimination and high resolution melting analysis
Syllabus
  • PCR and real-time PCR principles
  • Semiquantitative and quantitative PCR
  • PCR kinetics, end point vs. real-time PCR
  • Absolute and relative quantification, PCR efficiency
  • Template manipulation and quality, storage
  • Reverse transcription
  • Principles of fluorescence emmission and detection
  • Reporter fluorophores, quenchers, probe structure
  • Primer and probe design
  • General applications of real-time PCR
  • Experimental design, controls, reference genes, good laboratory practice
  • Data interpretation: absolute and relative quantification, SNP, high resolution melting analysis
  • Factors influencing PCR, normalisation, design of student's own experiments
  • MIQE Standards (Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines)
Literature
  • Real-time PCR. Edited by M. Tevfik Dorak. New York: Taylor & Francis, 2006, xxvi, 333. ISBN 041537734X. info
  • A-Z of quantitative PCR. Edited by Stephen A. Bustin. La Jolla, Calif.: International University Line, 2004, xxix, 882. ISBN 0963681788. info
Teaching methods
lectures
Assessment methods
Lectures
Final test
Language of instruction
Czech
Follow-Up Courses
Further comments (probably available only in Czech)
Study Materials
The course is taught annually.
The course is taught: in blocks.
Information on the extent and intensity of the course: v týdenním bloku.
The course is also listed under the following terms Autumn 2010 - only for the accreditation, Autumn 2008, Autumn 2009, Autumn 2011, Autumn 2011 - acreditation, Autumn 2012, Spring 2014, Spring 2015, Spring 2016, Spring 2017, spring 2018, Spring 2019, Spring 2020, Spring 2021, Spring 2022, Spring 2023, Spring 2024.
  • Enrolment Statistics (Autumn 2010, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2010/Bi9310