[Zbrafish] Re: sperm freezing for strain maintenance

Cecilia Moens via zbrafish%40net.bio.net (by cmoens from fhcrc.org)
Fri Oct 1 18:28:17 EST 2010


We freeze all of our mutant stocks and transgenic lines in house, according to the protocol published here: J Vis Exp. 2009 Jul 6;(29). The alternative is to submit your mutants to ZIRC, but ZIRC can only accept characterized mutants, so if you want to preserve mutants from a screen for possible future characterization, you'll have to do it yourself.

We try to freeze sperm from 10 males per line in duplicate vials in two different freezers, but even two or three vials is better than nothing. The sperm is good forever: we find that fertility is just as good for sperm frozen in 2001 as it is for sperm frozen last week. The variability in fertility is always huge: you'll see anywhere between 1% and 60% fertility for any given vial. But if you thaw into lots and lots of good eggs (up to 1000, pooled from multiple females) then even your 1% fertility vials will be recovered.

- Cecilia.

Cecilia B. Moens
Division of Basic Science
Fred Hutchinson Cancer Research Center
B2-152, 1100 Fairview Ave. N. 
Seattle, WA 98109-1024

office: (206) 667-5627
lab: (206) 667-5697
fax: (206) 667-3308

On Sep 28, 2010, at 10:03 AM, zbrafish-request from oat.bio.indiana.edu wrote:

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>   1. Sperm freezing for strain maintenance? (Burdine, Rebecca D)
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> Message: 1
> Date: Mon, 27 Sep 2010 13:34:33 -0400
> From: "Burdine, Rebecca D" <rburdine from Princeton.EDU>
> Subject: [Zbrafish] Sperm freezing for strain maintenance?
> To: <zbrafish from oat.bio.indiana.edu>
> Message-ID:
> 	<790DAC09623D0A47ACDB126E3C52D8B67A2F20 from MBCLUSTER.pu.win.princeton.edu>
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> 
> Hi,
> 
> 
> 
> I am wondering how many of you actually freeze sperm to maintain
> strains?  We are now in the position of having more strains that we
> really want to maintain, but we are reluctant to let some of them go in
> case we ever need them again.
> 
> 
> 
> In addition, I have already had one maintenance nightmare where a room
> was flooded with bleach and ozone (not compatible to fish health if you
> were wondering).  If something like that happened to my room with our
> mutants, I would be running a bookstore in upstate Vermont.
> 
> 
> 
> So...how reasonable is this to do?  How long do the stocks last and how
> often should they be tested/replenished?
> 
> 
> 
> 
> 
> Becky
> 
> 
> 
> ---------------------------------------------------
> 
> Rebecca D. Burdine, Ph.D.
> 
> Assistant Professor
> 
> Dept. of Molecular Biology
> 
> Princeton University
> 
> Washington Road Mof 433
> 
> Princeton, NJ 08544 
> 
> 
> 
> Phone: (609) 258-7515
> 
> Fax: (609) 258-6730
> 
> Email: rburdine from princeton.edu <mailto:rburdine from princeton.edu> 
> 
> Admin Assistant: Anna Schmedel (609) 258-5028
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