Protocols

Generation of transgenic mice with Yeast Artifial Chromosomes (YACs) and Bacterial Artificial Chromosomes (BACs)
 

  • Preparation of YAC-DNA for microinjection (PDF file). This method can also be applied for BAC-DNA. Download it here
  • PFGE conditions for the isolation of BAC DNA (PDF file). Download it here

    • Alternative method for producing YAC-DNA transgenic mice:

    Efficient generation of transgenic mice with intact yeast artificial chromosomes by intracytoplasmic sperm injection.
    Moreira PN, Giraldo P, Cozar P, Pozueta J, Jiménez A, Montoliu L, Gutiérrez-Adán A.
    Biol Reprod. 2004 Dec;71(6):1943-7. Epub 2004 Jul 30.  PDF
     

    Visit the Transgenic Animal Model Core of the University of Michigan for an additional protocol of BAC DNA preparation for microinjection

    Methods for preparing plasmid DNA for mouse oocyte microinjection
     

  • Preparation of plasmid DNA for microinjection, using glassmilk methods (RTF file, about 20 kb) Download it here
  • Preparation of plasmid DNA for microinjection, using NaCl gradient (RTF file, about 12 kb, also in Spanish!) Download it here
  • Preparation of plasmid DNA for microinjection [protocol from University of Michigan Transgenic Animal Model Core Facility ] (RTF file, about 12 kb) Download it here

    •    

    Preparing genomic DNA and analysing by PCR
     

  • Preparation of genomic DNA from mouse tail biopsies (RTF file, about  10 kb) Download it here
  • Mouse genotyping by PCR (RTF file, about 10 kb) Download it here

    •   

    Chi-Square Tests (observed/expected distribution of genotypes/phenotypes in mouse crosses involving 1, 2 or 3 unlinked loci)
     

  • Excel Workbook document (automatically calculates the Chi-Square value and the statistical significance of the observed cases) Download it here

    • Reference:

    Mendel: a simple excel workbook to compare the observed and expected distributions of genotypes/phenotypes in transgenic and knockout mouse crosses involving up to three unlinked loci by means of a χ(2) test.
    Montoliu L.
    Transgenic Res. 2012 Jun;21(3):677-81. Epub 2011 Aug 19. PDF



       
     

    Cryopreservation of sperm and mouse embryos (WEB page in "Transgenesis in Mammals")

    Back to initial page
     

    Lluís Montoliu (2011)