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Mouse Colony Management
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Standard Operating Procedure for Mouse Colony Management
Breeding Founders from Microinjection
- Founder mice breeding will start when they reach
- A male founder at six to eight weeks of age will
mate with two females.
- A female founder at four to six weeks of age will mate
with one male.
- The GTTF will use wild type littermates generated
from the same microinjection for mating or mice from
vendors if requested.
- Tail snips or ear notches will be given to the
investigator for genotyping analysis.
- F1 mice from Indels or Ki/+ founder mice from CRISPR
microinjection should be sequenced to confirm gene
- In order to preserve the transgenic line the CMGM
could retain two Tg/+ males and two Tg/+ females for
- The remaining mice will be available to the
investigator for experimental use or to characterize the
Expansion of Transgenic Line (Establishing the Line)
- Heterozygous breeding will be set up according to the
investigators' requests and needs. A breeding cage can
contain one male and two female mice. The females will be
separated when they are obviously pregnant.
- The CMGM will always keep one cage of breeding age
transgenic males. All other mice of the same line will
be available for experimental use.
- Upon the request of an investigator, the wildtype mice
will be culled if they are not to be used for
experimental purposes. These wildtype mice can be used
for breeding within the same line. Individual
investigators may choose to purchase wildtype mice for
breeding at their own cost.
Establishing Homozygous Transgenic Mouse Lines
- Breeding to Homozygousity
- Tg/+ X Tg/+ mating will be set up, two pair for each
- Litters will be screened for homozygous transgenic mice
by the investigator.
- Test Homozygousity
- Possible homozygous mice will mate with wildtype mice
to test for true homozygosity.
- To reduce the cost CD-1 or other wildtype mice can be
used for these test matings. Investigators can also
order wildtype mice of the same strain at their own
- The pups from these crosses will be sacrificed at
weaning and tails snips will be screened. Litters from
these crosses can be kept at the request of the
- A mouse with more than ten offspring that are all Tg/+
will be considered as homozygous. If any of the
offspring are +/+ the parent is a heterozygous mouse and
will be euthanized as long as the line has sufficient
- Breeding Homozygous Mice
- When a pair of mice is identified as homozygous, the
breeding will be set up to generate homozygous mice.
- The GTTF will keep one cage of males and one cage of
females. The reserves will be maintained at breeding
- All other mice generated from these crosses will be
available for experimental use.
Chimeric Mice Breeding
- A chimeric mouse will mate with one wildtype CD-1
female at six weeks of age to test germline transmission.
- When germline transmission chimeras are identified,
the breeding will be set up according to the
- To generate 129Sv/B6/CD-1 mice: A 129Sv/B6 chimeric male will mate with CD-1 female mice.
- To generate 129Sv/B6 floxed mice: A 129Sv/B6 chimeric
male will mate with B6 or 129 FLP females.
- To generate 129Sv/B6 knockout mice: A 129Sv/B6 chimeric
male will mate with B6 Hprtcre females.
- To generate congenic knockout mice: chimeric male
will mate with inbred strain mice chosen by the
- CD-1 females and either FLP or Hprtcre females (investigtor's
choice) used in breeding will be provided by the
GTTF. Additional females are purchased at
the investigator's expense.