International Institute of Molecular and Cell Biology 2 articles published in JoVE Genetics In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity Krzysztof J. Skowronek1, Matthias Bochtler1 1International Institute of Molecular and Cell Biology, Warsaw (IIMCB) Restriction endonucleases with new sequence specificity can be developed from enzymes recognizing a partially degenerate sequence. Here we provide a detailed protocol that we successfully used to alter the sequence specificity of NlaIV enzyme. Key ingredients of the protocol are the in vitro compartmentalization of the transcription/translation reaction and selection of variants with new sequence specificities. Biology A Protocol for Laboratory Housing of Turquoise Killifish (Nothobranchius furzeri) Joanna Dodzian1,3, Sam Kean1, Jens Seidel1, Dario Riccardo Valenzano1,2 1Max Planck Institute for the Biology of Ageing, 2CECAD, University of Cologne, 3International Institute of Molecular and Cell Biology in Warsaw Laboratory housing of turquoise killifish can be scaled up to house and efficiently raise thousands of individual fish in a centralized water filtration system, employing the same infrastructure used for standard zebrafish facilities. Here we detail a list of standardized procedures that allow efficient killifish maintenance.
