Sign-up Guide
Before signing up for a course please take a look at "The current state of free spots" to verify that the course you chose has a spot for you (link below). Each course has an individual minimum and a maximum number of attendees. The minimal number of attendees to open the course is listed in the course label and is also indicated by green color in the course column. After you pick a course with free spot, click on the "Course Signup" button that will redirect you to a form, where you fill out your name and email address. You can verify that you have been signed up by checking "The current state of free spots" again. Your name should appear in an appropriate column within a few minutes of signing up.
Please note that if your name appears in the red cell it means that you exceed the capacity of a course and you need to sign up to a different course.
The current state of free spots
Course Descriptions
Course 1
Guarantor: Adam Bajgar
Name of the technique: qPCR expression analysis
Responsible person name: Gabriela Krejčová
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 12 hours
Short description: Practice will consist of a selection of concrete gene, work with analysis and design of
qPCR primers. The tissue samples will be used for RNA isolation and reverse transcription. qPCR by using
SYBR green will be followed by data analysis by method double delta.
 Course 1 Sign Up |
*** DISCONTINUED ***
Course 2
Guarantor: Tomáš Doležal
Name of the technique: Advanced Drosophila genetics – generation and analysis of mitotic clones
Responsible person name: Tomáš Doležal
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 3 days
Short description: Theoretical learning of advanced Drosophila genetics and crosses for mitotic clone
generation, fly handling and phenotyping under the microscope, larval dissection and microscopy
sample preparation, confocal microscopy analysis of clones.
Course 3
Guarantor: David Doležel
Name of the technique: Systemic RNA interference in insects
Responsible person name: David Doležel
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 4-5 afternoons
Short description:
Day 1: PCR to prepare a template for RNA transcription (theory during the PCR reaction),
Day 2: Product purification + in vitro transcription (O/N)
Day 3: dsRNA annealing & Injection into P. apterus
Day 4: Phenotype assessment – 2 weeks later – 2hrs during the afternoon
| Course 3 Sign Up |
*** DISCONTINUED ***
Course 4
Guarantor: Alexander W. Bruce
Name of the technique: Microinjections and immuno-fluorescence staining of mouse embryos
Responsible person name: Pablo Bora (Ph.D. student)
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 4-5 days
Short description: Microinjections (of mRNAs, short RNAs or proteins) are a common technique used for
the gene down-regulation or overexpression in mammalian oocytes and embryos. We will inject 1 cell of
mouse 2-cell stage embryos with fluorescently-tagged mRNA, embryos will be cultured until the early
blastocyst stage and then they will be fixed and immuno-stained for a marker protein of trophoblast cell
lineage (future placenta). Confocal microscopy will be used to visualise injected fluorescent protein and
immuno-stained trophoblast cell lineage marker, differentiating between the progeny of injected and
the non-injected cell at the 2-cell stage, as well as between the cells that will contribute to the placenta
(presence of trophoblast marker protein) and the embryo itself (absence of trophoblast marker protein).
Course 5
Guarantor: Petr Nguyen
Name of the technique: Fluorescence in situ hybridization
Responsible person name: Petr Nguyen
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 3 days
Short description: Students will learn how to make chromosome preparations, construct fluorescently
labelled probes and hybridize them to chromosomes. Probes can be constructed from a whole genome,
clone from the genomic library, repetitive sequence or single-copy gene.
 Detailed protocol |
Course 5 Sign Up |
Course 6
Guarantor: Tomáš Korytář
Name of the technique: Flow cytometry and cell sorting
Responsible person name: Tomáš Korytář
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 3 days
Short description: In this course, students will learn the basic principles of flow cytometry and its
applications in life sciences on routinely used assays. The key aim is to provide students with the
knowledge necessary for performing any flow cytometry application from the design of the experiment,
design of antibody panels, sample acquisition and subsequent analysis and data presentation.
The course is open only for students planning to use flow cytometry and cell sorting in their research.
| Detailed protocol |
Course 6 Sign Up |
Course 7
Guarantor: Zdeněk Paris
Name of the technique: Denaturing polyacrylamide gel electrophoresis and radioactive detection of small non-coding RNAs by Northern blot
Responsible person name: Zdeněk Paris
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 4 days
Short description: Urea PAGE or denaturing urea polyacrylamide gel electrophoresis employs 6-8 M
urea, which denatures secondary RNA structures and is used for their separation in a polyacrylamide
gel matrix based on the molecular weight. Fragments between 2 to 500 bases, with length differences
as small as a single nucleotide, can be separated using this method. The migration of the sample is
dependent on the chosen acrylamide concentration. A higher percentage of polyacrylamide resolves lower
molecular weight fragments. The combination of urea and temperatures of 45-55 °C during the gel run
allows for the separation of unstructured RNA molecules. After the PAGE, the gel is transferred to a nitrocellulose
membrane and hybridized with a P32 radiolabeled oligonucleotide probe to visualize the specific RNA
species using the Phosphoimager.
| Course 7 Sign Up |
Course 8
Guarantor: Roman Sobotka
Name of the technique: 2D native/SDS electrophoresis combined with immunodetection
Responsible person name: Petra Skotnicová
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 2.5 days
Short description: During the course, students will learn to set a large electrophoretic system (Bio-Rad
Protean xi cell), to pour and run blue-native and clear-native 8-14% gradient gels with samples of
membrane protein complexes. Gel strips from the first dimension will be separated overnight on a 12-
20% SDS gel, stained with SYPRO Orange, blotted onto PVDF membrane and probed with selected
antibodies.
| Course 8 Sign Up |
*** DISCONTINUED ***
Course 9
Guarantor: Roman Sobotka
Name of the technique: Expression and purification of His-tagged proteins
Responsible person name: Petra Skotnicová
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 2 days
Short description: Students will learn the transformation of E. coli by electroporation, cultivation of E. coli
cells, preparation cell lysate by sonication, purification of His-tagged fluorescent proteins on a nickel
column, analysis of purified proteins on a small SDS gel, Coomassie stain.
Course 10
Guarantor: Hana Sehadová
Name of the technique: Classical histological techniques and light microscopy
Responsible person name: Hana Sehadová, Markéta Vrchotová
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it., This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 2 days
Short description: 1. Preparation of paraffin sections of insect tissues and blood smear, 2. Mallory and
Giemsa staining, 3. Observation of obtained samples in the light microscope.
| Course 10 Sign Up |
Course 11
Guarantor: Tomáš Doležal
Name of the technique: Fluorescent labeling of the Drosophila central nervous system
Responsible person name: Ellen McMullen
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 3 days
Short description: Students will dissect Drosophila larval brains and perform multicolored labelling of neurons
by immune staining (Brainbow). We will use confocal microscopy to image these fixed samples as well as
monitoring glucose dynamics in the blood-brain barrier of live brains expressing a genetically encoded glucose sensor (FRET).
| Course 11 Sign Up |
Course 12
Guarantor: Eva Doleželová, Alena Zíková
Name of the technique: Measurement of mitochondrial respiration and mitochondrial
membrane potential by High-Resolution Respirometry (HRR) using the O2k-
FluoRespirometer
Responsible person name: Michaela Kunzová
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 1 day
Short description: The students will get a theoretical introduction into the High-
Resolution Respirometry, the principle and what it is used for. Then during the practical
session, they will learn how to properly operate the O2k-FluoRespirometer and measure
respiration in intact and digitonin permeabilized cells adding different substrates and
inhibitors. Taking advantage of the new O2k-TPP+ ISE-Module, the students will be able
to simultaneously with the respiration monitor also the mitochondrial membrane potential
in digitonin permeabilised cells by using Safranine O or TMRM stain.
| Detailed protocol |
Course 12 Sign Up |
Course 13
Guarantor: Marek Jindra
Name of the technique: Western blot
Responsible person name: Marek Jindra
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 2 days
Short description: In this hands-on course students will analyze the expression of specific proteins of
interest either in insect tissues or in cultured cells. They will prepare tissue/cell extracts, prepare and
run denaturing polyacrylamide gel electrophoresis, and detect proteins after immunoblotting using
antibodies and chemiluminescence.
| Course 13 Sign Up |
Course 14
Guarantor: Petr Nguyen
Name of the technique: NGS data processing
Responsible person name: Petr Nguyen
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 3 days
Short description: Students will learn basic processing of NGS data
Detailed protocol: Bash basics; cluster computing within the Metacentrum infrastructure; data download; quality check
and filtering; de novo assembly and read mapping.
| Course 14 Sign Up |
Course 15
Guarantor: Lenka Gahurová, Iva Mozgová
Name of the technique: RNA-seq library preparation and data analysis
Responsible person name: Iva Mozgová, Lenka Gahurová
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it., This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 2 days
Short description: RNA sequencing is used to study changes in RNA expression at the genome-wide level
either at different time point or at different conditions. This course will teach you: 1. How to find the
best approach and the best kit for RNA-seq library preparation of your samples 2. How to find the best
sequencing strategy 3. How to check the quality of your library 4. How to check the quality of your data
5. How to analyse your data to identify differentially expressed genes.
| Detailed protocol |
Course 15 Sign Up |
Course 16
Guarantor: Michal Žurovec
Name of the technique: Transfection of insect cell lines
Responsible person name: Václav Brož
Responsible person email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Duration of the course: 3 days
Short description: We will show students different types of insect cell cultures, teach them DNA
transfection to the cells and detection of marker expression.
| Detailed protocol |
Course 16 Sign Up |
