Bi6721c Special Methods of Microorganisms Analysis I. - practical course

Faculty of Science
Spring 2022
Extent and Intensity
0/3/0. 3 credit(s). Type of Completion: z (credit).
Teacher(s)
doc. Mgr. Pavel Dvořák, Ph.D. (seminar tutor)
Mgr. Barbora Burýšková (seminar tutor)
Mgr. Barbora Popelářová (seminar tutor)
Guaranteed by
doc. Mgr. Pavel Dvořák, Ph.D.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: Ing. Jiřina Kučerová, Ph.D.
Supplier department: Department of Experimental Biology – Biology Section – Faculty of Science
Timetable
Mon 13:00–18:50 E25/119
Prerequisites
NOW( Bi6721 Spec. Meth. Microorg. Anal. )
Prerequisite for registration of the lab practice is parallel enrollment in the course Special Methods of Microorganisms Analysis (Bi6721).
Course Enrolment Limitations
The course is also offered to the students of the fields other than those the course is directly associated with.
The capacity limit for the course is 10 student(s).
Current registration and enrolment status: enrolled: 0/10, only registered: 0/10, only registered with preference (fields directly associated with the programme): 0/10
fields of study / plans the course is directly associated with
there are 6 fields of study the course is directly associated with, display
Course objectives
The aim of the practical course is to acquaint students with the method of isolation of plasmid and genomic DNA of bacteria using a kit, with the design of oligonucleotide primers and preparation of polymerase chain reaction (end-point and quantitative real-time PCR), with the method of control gel electrophoresis, sequence analysis, and with the purification of the foreign protein from the host bacterium by affinity chromatography.
Learning outcomes
At the end of this course, the students should be able to use practically the microbial DNA isolation protocol, the method of polymerase chain reaction, sequence analysis, and the gel electrophoresis method.
Syllabus
  • 1. Preparation of bacterial lysates from cultures of Pseudomonas putida and Escherichia coli.
  • 2. Amplification of four specific genes.
  • 3. Gel electrophoresis of the amplification, purification of the PCR products.
  • 4. Electrophoresis of the purification PCR products, quantification and preparation of the sample for the sending and DNA sequencing.
  • 5. In silico analysis of the DNA sequencing.
  • 6. Isolation of the genomic DNA - bacterial culture Escherichia coli.
  • 7. Estimation of the concentration and purity of the DNA spectrophotometrically.
  • 8. Indicative estimation of the concentration and the purity of the DNA spectrofotometrically.
  • 9. Polymerase chain reaction by system end-point, gel electrophoresis.
  • 10. Real-time PCR with DNA sample from cytomegalovirus.
  • 11. Purification of a recombinant protein from Escherichia coli using affinity chromatography
Literature
  • Green, M.R., Sambrook, F. Molecular Cloning. A Laboratory Manual. 4th ed. Cold Spring Harbor Laboratory Press. 2012

    ​Kopecká, J., Bartoš, M. Speciální metody analýzy mikroorganizmů I, návody ke cvičení​, 2014

Teaching methods
The practice takes place once in 14 days. Students work individually and process their own sample. The results of experiments have to be summarized and evaluated in protocols. The protocols are prepared in accordance (information v  the first practice and lecture). In the protocols students describe, compare, analyze, evaluated and discuss own results.
Assessment methods
The course is closed by a course-unit credit. The students obtain their credits for active attendance at the course, for the preparation of protocols, and for a brief discussion with the lecture about the prepared protocols.
Language of instruction
Czech
Follow-Up Courses
Further comments (probably available only in Czech)
Study Materials
The course is taught annually.
General note: Předmět se doporučuje zapsat ve 2. semestru společně s přednáškou, případně pak ve 4. semestru.
Information on course enrolment limitations: Přednost mají posluchači specializace mikrobiologie.
The course is also listed under the following terms Spring 2008 - for the purpose of the accreditation, Spring 2011 - only for the accreditation, Spring 2003, Spring 2004, Spring 2005, Spring 2006, Spring 2007, Spring 2008, Spring 2009, Spring 2010, Spring 2011, spring 2012 - acreditation, Spring 2014, Spring 2016, Spring 2017, spring 2018, Spring 2019, Spring 2020, Spring 2021, Spring 2024, Spring 2025.
  • Enrolment Statistics (Spring 2022, recent)
  • Permalink: https://is.muni.cz/course/sci/spring2022/Bi6721c