PřF:Bi9310 Real-time PCR - Course Information

Bi9310 Real-time polymerase chain reaction

Extent and Intensity

2/0/0. 2 credit(s) (plus extra credits for completion). Recommended Type of Completion: zk (examination). Other types of completion: k (colloquium).

Teacher(s)

doc. RNDr. Sabina Ševčíková, Ph.D. (lecturer)

Guaranteed by

doc. RNDr. Sabina Ševčíková, Ph.D.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: doc. RNDr. Sabina Ševčíková, Ph.D.
Supplier department: Department of Experimental Biology – Biology Section – Faculty of Science

Prerequisites

Bi4035 Molecular biology - lab.pract. || Bi4020c Molecular biology - practice
General knowledge in the field of Molecular Biology.

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

there are 9 fields of study the course is directly associated with, display

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 allelic discrimination and high resolution melting analysis

Learning outcomes

At the end of this course, the student will understand the principles of PCR and real-time PCR; he/she will be capable of designing and performing a real-time PCR experiment, including all controls; he/she will have general knowledge on the structure of fluorophores and probes used in this technique and skills to analyze data from absolute and relative quantification, SNP analysis 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 emission 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

• Bi9310c Real-time polymerase chain reaction – practice

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.

• Enrolment Statistics (Spring 2023, recent)
  
• Permalink: https://is.muni.cz/course/sci/spring2023/Bi9310