zebrafish e-book

1、THE ZEBRAFISH BOOKA Guide for the Laboratory Use of Zebrafish Danio (Brachydanio) rerioby Monte Westerfield, Institute of Neuroscience, University of Oregon3Preparing Embryos for Cell Culture.............................

2、...................................................................1590 Ca2+ - Mechanical Dissociation...................................................................................................159Protease Dissoci

3、ation .......................................................................................................................159Dissociation of Embryos by Dissection ......................................................

4、....................................160Using Coated Glass Coverslips for Cell Culture...........................................................................160Recipes:.................................................

5、..............................................................................................160CHAPTER 7 - GENETIC METHODS ...............................................................................................

6、......162Conventions for Naming Zebrafish Genes ....................................................................................162Suggested Guidelines for Maintaining Mutant stocks...................................

7、...............................164Overview of Methods for Parthenogenesis....................................................................................166Production of Haploid Embryos...............................

8、........................................................................169Production of Homozygous diploid embryos.................................................................................170DELAYED IN VITRO FERT

9、ILIZATION USING COHO SALMON OVARIAN FLUID..............175PRODUCTION OF ANDROGENETIC HAPLOIDS AND DIPLOIDS.........................................178Freezing Sperm........................................................

10、..........................................................................182Thawing and Using Frozen Sperm for In Vitro Fertilization ........................................................183Fin Amputations ..........

11、.......................................................................................................................184Gamma Ray Mutagenesis.............................................................................

12、...................................184Chemical Mutagenesis......................................................................................................................185Testosterone Treatment to Produce Males .

13、..................................................................................187ZEBRAFISH STRAINS .....................................................................................................................

14、188CHAPTER 8 - HISTOLOGICAL METHODS .........................................................................................189Preparing subbed slides for sections........................................................

15、......................................189OCT Embedding for Cryostat Sectioning of Embryos or Larvae ...............................................189Agar Embedding for Cryostat Sectioning of Embryos or Larvae ..........

16、.....................................190Gelatin Embedding for Vibratome Sectioning of Embryos or Larvae........................................191Staining Sections with Antibodies Using PAP...............................

17、................................................192Staining Whole-Mount Embryos with PAP ....................................................................................193Solutions for Antibody Staining Protocols...

18、...................................................................................195Whole-Mount Staining of Biotin-Dextran Injected Zebrafish Embryos......................................197Photoconversion of Fluoresce

19、ntly Labeled Profiles for EM ........................................................198Chromosome Spreads..................................................................................................................

20、....200Gentle Fixation by Freeze Substitution ..........................................................................................201The following is a list of available antibodies at ZIRC: ........................

21、........................................203Chapter 9 - Molecular Methods ...............................................................................................................209Extraction of Proteins from Zebra

22、fish Embryos for SDS-Gel Analysis......................................209Western Blots of Zebrafish Embryos..............................................................................................209Preparation of

23、Genomic DNA ..........................................................................................................213Preparation of Genomic DNA for Southern Analysis ..................................................

24、.................215Rapid Isolation of RNA from Zebrafish...........................................................................................216Extraction and Purification of RNA from Zebrafish Embryos............

25、..........................................216Preparation of High Quality RNA from Zebrafish Embryos.........................................................217Whole-Mount in situ Hybridization ...........................