1. Pages 2 to 7: Playing cards for Memory and Total recall Pages 2 to 4: keyword-cards Pages 5 to 7: keyword-cards with corresponding explanation-cards Page 8: Playing cards for Organogram

2. Restriction Restriction Star fragment endonuclease activity Isoschizo- Gel mer electrophoresis Slot Growth Ethidium Electrophoretic rate bromide mobility Double Origin of Selectable digestion replication marker pBR 322 MCS Polylinker Complex  -Galacto- High copy media sidase plasmid Low copy Defined DNA plasmid media ligases Competent Escherichia pUC 19 cells coli Electro- Ligation OD poration

3. EcoR I lac-Operon Plasmid Cloning Electropho- Base vector resis pair Antibiotic Blunt end Sticky end resistance enzymes enzymes Restriction Endo- map nucleases Palindrome DNA Recognition Target band site sequence Restriction Compatible enzyme Ampicillin ends Electropho- Restriction Salt resis marker buffer competent lacZ gene ORI Heat shock Bromo- Xylen- Type I phenol blue xyanol enzyme

4. Hybridi- Probe Label sation Denatura- Southern RBS tion Blot Target Promoter Terminator sequence Expression Southern Western vector Blot Blot Agarose PCR Primer Annealing Synthesis Cycle Cassette

5. DNA modifying enzymes that recognise specific motifs of double stranded DNA and cleave it into fragments of different lengths type II restriction enzyme: recognises a specific palindrome sequence motif (GAATTC) sequence motif with an axis of rotational symmetry, the sequence in 5`  3`-direction is equal on the both strands small rings of double stranded extrachromosomal and independently reproduced DNA, natural types are important for antibiotic resistance and sexual reproduction of bacteria vehicle of genetic engineering shuttle that allows reintroducing of foreign DNA into a host organism Endonucleases Palindrome Plasmids EcoRI Vector

6. bacterial plasmids with features for reintroducing of small foreign DNA-fragments into host organisms with the purpose of producing significant amounts of recombinant DNA Cloning vector diagram showing sites of cleavage of a DNA molecule, for example of a plasmid vector Restriction map Cleavage of DNA with this type of endonucleases creates a short complementary single stranded overhang in each cleaved end Sticky end enzymes Blunt end enzymes Endonucleases that form no 5’ or 3’ overhangs after cleavage Competent cells Bacterial cells which are able to incorporate foreign DNA

7. lac-Operon Bacterial expression unit of genes for lactose utilisation Agarose gel electrophoresis Method that separates DNA fragments in an electric field as a function of their lengths Polylinker Staged recognition sites of different restriction enzymes, a feature of vectors for variable insertion of foreign DNA Isoschizomer Restriction enzymes that recognise the same target sequence Ethidium bromide (EtBr) Stain that intercalates into double-stranded DNA. Under UV-light the complex of DNA and the stain fluoresces pink

8. DNA-cloning Restriction of vector and insert Ligation TransformationVerification Restriction with the same enzyme(s) Restriction with different enzymes Sticky end ligation is easier 1 hour, 37 °C Blunt end ligation is more difficult 16 hours, 4 °C Competent cells plus ligation reaction Picking of transformants Heat shock Over night cultivation with antibiotics Cultivation without antibiotics Plasmid DNA isolation Cultivation on plates with antibiotics Digestion or PCR Agarose gel electrophoresis Result