Services
About the services
Jump to: Making a fish line — Molecular cloning — Phenotyping — Genotyping — IVF — Cryopreservation
Making a fish line
CRISPR-Cas9/Tol2/phiC31: Generation of a transgenic (Tol2/pIGLET) or mutant (frameshift/loss of function) fish line through F2 generation plus early phenotyping.
Homology-directed repair: Generation of a transgenic fish line with a precise edit (e.g. a specific amino acid substitution).
Molecular cloning
One-step LR gateway reaction: Covers assembly of Gateway reactions from existing Entry clones.
Two-step cloning (pENTR5’ or pME + LR gateway reaction): Covers assembly of Gateway reactions requiring generation of Entry clones (e.g. BP + LR reactions).
Phenotyping
Group 1: plasmid/mRNA/CRISPR-Cas9 injection F0: Phenotypic analysis following an injection of plasmid DNA, mRNA, or CRISPR/Cas9 nucleoproteins in the resulting F0 fish; generally one control, one gene of interest, one analysis.
Group 2: immunohistochemistry: This encompasses antibody staining or in situ hybridization to localize proteins or transcripts of interest.
Imaging: Routine confocal or brightfield imaging of lines or tissue samples already generated by some other method.
Behavioral phenotyping: Analysis of zebrafish behavior (generally at 5 days post-fertilization) in auditory-evoked escape or circadian behavioral paradigms (although other paradigms are possible, please contact us to discuss further).
Monitoring: Any tanks which require management by Core staff after completion of Making a Fish Line projects are charged a nominal monthly fee to cover administrative costs.
Genotyping
Genotyping of adults or embryos for any lines which are NOT en route to F2 as part of a Making a Fish Line project; discounts are available once a suitable assay has been developed.
In vitro fertilization
Isolation and combination of zebrafish gametes (usually this is employed to rescue or propagate lines that have become too old or sick to breed naturally).
Cryopreservation
Isolation and sperm freezing from ≥10 males for long-term storage and later revival; there is no maintenance fee for storage.
Available to Yale researchers & external researchers
Training & services
Basic training: Lab staff who wish to learn to work with zebrafish receive one or more training sessions (each session is billed separately) of 2+ hours with core staff. This generally covers fish husbandry and microinjection, although we are able to train users in diverse techniques as needed.
Consultation: Meeting with core staff to discuss new or ongoing projects; the first meeting on any given project will not be charged.
Protocol, grant, & manuscript assistance: We assist with writing an IACUC protocol, grant application, or manuscript to which we have contributed data. The fee here is charged once per document.
Using Zebrafish Core equipment unassisted: Trained users working with core equipment are assessed a small hourly fee to cover repair and maintenance costs; this does not cover fees associated with use of the LightCycler or behavioral unit.
Staff service: Core technical staff can be employed for contract injection (or other) work involving provided constructs or reagents; this does not cover phenotyping or other downstream analyses.