Molecular Techniques in Biology 320-ERS-1MTE
Laboratorium (LAB) Rok akademicki 2020/21

1. Carson S. 2019. Molecular Biology Techniques. A Classroom Laboratory Manual. Elsevier Science Publishing Co Inc

2. Rapley R. 2014. Molecular Biology and Biotechnology. Royal Society Of Chemistry

1. Student explains the principles of particular methods of Molecular biology.

2. Student characterizes the advantages and disadvantages of particular methods of Molecular biology and knows their applications.

3. Student distinguishes between the methods of DNA fragment size analysis and DNA sequence analysis and is able to apply them in practice.

4. Student is aware of the research and application possibilities offered by the tools of Molecular biology.

5. Using an electrophoresis apparatus, a DNA thermocycler and a sequencer, student performs basic molecular analyzes to determine the genotype of the organism under study.

6. Student is able to work in a team and independently while performing laboratory and computer analyzes in the field of Molecular biology.

A short, written answer to the question asked.

1. DNA isolation and gel electrophoresis.

a) Main rules of work in Molecular Biology Laboratory.

b) Pipetting practice.

c) DNA extraction from animals tissues.

d) Gel electrophoresis.

2. PCR - sex identification of mammals.

a) PCR-MULTIPLEX

b) Sex identification of mammals.

c) Gel electrophoresis.

3. Identification of polymorphisms in mitochondrial control region by PCR-RFLP.

a) Digestion of PCR products (fragment of mitochondrial ctr of common vole Microtus arvalis) with HaeIII (BsuI) enzyme (RFLP).

b) Simulation of digestion – NEBcutter V2.0

c) Gel electrophoresis.

4. Molecular species identification – part 1.

a) Clean-up of PCR products.

b) Sequencing reaction.

5. Molecular species identification – part 2.

a) Purification of sequencing reaction products with the ExTerminator kit (A&A Biotechnology).

b) Separation of sequencing products on a 3130 Genetic Analyzer (Applied Biosystems).

c) Results interpretation: BLAST, NCBI website.

6. Molecular species identification – part 3.

a) Working with the sequences obtained - ChromasLite and BioEdit programs.

b) Results interpretation - BLAST, NCBI website.

c) Constructing phylogenetic trees - MEGA program.

1) laboratory work with Instructions