Transcript ppt
Playing cards for Memory and Total recall Pages 2 to 4: keyword-cards Pages 5 to 7: keyword-cards with corresponding explanation-cards Restriction Restriction fragment endonuclease IsoschizoGel mer electrophoresis Growth rate Double digestion Ethidium bromide Origin of replication Star activity Slot Electrophoretic mobility Selectable marker pBR 322 MCS Polylinker Complex media -Galactosidase High copy plasmid Low copy plasmid Defined media DNA ligases Competent Escherichia cells coli Ligation OD pUC 19 Electroporation EcoR I lac-Operon Cloning vector Electrophoresis Antibiotic resistance Restriction map Blunt end enzymes Endonucleases DNA band Restriction enzyme Recognition site Ampicillin Plasmid Base pair Sticky end enzymes Palindrome Target sequence Compatible ends Electropho- Restriction resis marker buffer Salt competent lacZ gene ORI Heat shock Bromophenol blue Xylenxyanol Type I enzyme Hybridisation Probe Label Denaturation Southern Blot Target sequence Expression vector Promoter Terminator Southern Blot Western Blot Agarose PCR Primer Annealing Synthesis Cycle RBS Cassette DNA modifying enzymes that recognise specific motifs of double stranded DNA and cleave it into fragments of different lengths sequence motif with an axis of rotational symmetry, the sequence in 5`3`-direction is equal on the both strands Endonucleases Palindrome vehicle of genetic engineering shuttle that allows reintroducing of foreign DNA into a host organism small rings of double stranded extrachromosomal and independently reproduced DNA, natural types are important for antibiotic resistance and sexual reproduction of bacteria type II restriction enzyme: recognises a specific palindrome sequence motif (GAATTC) Vector Plasmids EcoRI 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 Endonucleases that form no 5’ or 3’ overhangs after cleavage Blunt end enzymes Bacterial cells which are able to incorporate foreign DNA Competent cells Bacterial expression unit of genes for lactose utilisation lac-Operon Method that separates DNA fragments in an electric field as a function of their lengths Agarose gel electrophoresis Restriction enzymes that recognise the same target sequence Isoschizomer Staged recognition sites of different restriction enzymes, a feature of vectors for variable insertion of foreign DNA Polylinker Stain that intercalates into double-stranded DNA. Under UV-light the complex of DNA and the stain fluoresces pink Ethidium bromide (EtBr)