ZIRC Protocols

These protocols are presented in both PDF () and text-only versions. The PDF version is recommended. If you do not already have the latest PDF reader, it may be downloaded from Adobe (new window).

Antibody Labeling | Approved IACUC Animal Use Protocols | Cryopreservation and In Vitro Fertilization (IVF) | Egg Bleaching | Fish Feeding | Fish Nursery | Genotyping | ZIRC Health Monitoring SOPs | Labels

Antibody Labeling
HIAR pdf
Heat Induced Antigen Retrieval (HIAR) This protocol describes a pretreatment of tissues for optimal antigen target preservation that is used for some monoclonal antibodies.
Wholemount Antibody & ABC reaction pdf
This is a standard labeling protocol for antibody labeling and additional ABC staining.
Wholemount Antibody Labeling pdf
This is a standard labeling protocol derived from the Zebrafish Book (Westerfield, 2000).
Approved IACUC Animal Use Protocols
ZIRC pdf
This is the approved animal use protocol for the Zebrafish International Resource Center in Eugene, Oregon, USA.
Amacher Lab pdf
This is the approved animal use protocol for the Amacher lab at the University of California at Berkeley, located in Berkeley, California, USA.
Burgess Lab pdf
This is the approved animal use protocol for the Burgess lab at the National Institutes of Health, located in Bethesda, Maryland, USA.
Cryopreservation and In Vitro Fertilization (IVF)
Current Protocols
ZIRC E400/RMMB Sperm Cryopreservation & IVF Protocol by Matthews et al. V.1.2_2017_07_18 pdf
A step-by-step description of ZIRC's current cryopreservation & IVF protocols
Calibration Curve and Sperm Density Calculator xls
Sperm Density Calculator for use with Nanodrop Spectrophotometer concentration determination.
Previous Protocols
Cryopreservation of zebra fish spermatozoa using methanol by Harvey et al. 1982 pdf
The Canadian Journal of Zoology has kindly permitted us to provide a downloadable document of the original article by Harvey et al., (1982) that describes cryopreservation for zebrafish. All copyrights remain with the Canadian Journal of Zoology. You can download the article for your own use. Please inform yourself here about copyright issues.
Sperm Cryopreservation by Walker and Streisinger
The laboratory protocol that was adapted from Harvey et al. by Walker and Streisinger can be found in the Zebrafish Book, Chapter 7.
Sperm Cryopreservation by Draper and Moens. 2004
View Draper and Moens' complete article and protocol video (A high-throughput method for zebrafish sperm cryopreservation and in vitro fertilization) on the Fred Hutchinson Cancer Research Center web site. This is the protocol on which we base our procedures here at ZIRC (see 4, below).
Cryopreservation and In Vitro Fertilization at the ZIRC by Carmichael et al. 2009 pdf
This review provides a step-by-step description of the Draper and Moens method (see 3, above) as it is currently applied at the ZIRC. In addition, we include detailed notes with background, advice, and additional information. The authors, and ZIRC in general, wish to express their gratitude to Cecilia and Bruce for teaching us the protocol and for their continued advice and support.
Egg Bleaching
Egg Bleaching pdf
Fish Feeding
Dry Food
Dry Food Recipes pdf
Live Food
Artemia Decapsulation pdf
Decapsulated Artemia Hatching pdf
Non-Decapsulated Artemia Hatching pdf
ZIRC Paramecia Instructions pdf
Schedules and Suppliers
Feeding Schedule - weekday and weekend pdf
Food Source Information pdf
Fish Nursery
E2 Solution pdf
Basic Nursery Instructions pdf
Detailed Nursery and Food Instructions pdf
Nursery Protocol, Siphon Method pdf
A. Read this first!                                                       pdf
B. Overview of Genotyping Assays at ZIRC pdf
C. Designing and Handling of PCR Primers pdf
D. PCR Sample Preparation pdf
E. General PCR Protocol pdf
F. Restriction Enzyme Digestion pdf
G. Gel Electrophoresis pdf
H. Controls pdf
Files A. - H. Combined pdf
I. Find and Download Line Specific
   Genotyping Protocols  here
We provide all protocols we use at ZIRC online as soon
as possible. If you see a fish line that has no protocol
associated with it on our pages, it means that it probably
does not exist. Please be reminded that we are unable to
develop and provide specific PCR protocols upon request.
ZIRC Health Monitoring SOPs
Sentinel Fish Program pdf
Daily Monitoring of Fish Morbidity and Mortality SOP pdf
Fixing Zebrafish for Histopathology pdf
ZIRC PCR protocol for P. neurophilia pdf
ZIRC Label Info pdf