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If no litters are produced after one month you should replace the mice that you are mating with your founder.It is possible that the transgenic founder may be infertile due to consequences of transgene expression or unknown reasons. a) C57BL/6 is a standard inbred strain, commonly used in transgenic breeding b) mate 6 - 8 week-old mice for best reproductive performance i. replace females after 6 litters or when they are 6 months old c) mate a founder male with 2 females to get 2 litters in close succession d) mate a founder female with 1 male e) mice usually mate again on the day the female gives birth, resulting in a second litter 3 weeks after the first.You may be lucky get germline transmission at low efficiency (defined as 1 agouti pup in 200 pups born).separate the fighting mice, house them 1 per cage if necessary ii. males may fight in the following circumstances: male is placed in a cage containing other male(s) male is separated at weaning and then reunited with male littermates male is weaned into a cage that contains males from another litter males are aggressive and may begin to fight for no apparent reason adult male attacks immature female when female is placed in male's cage 6. house males and females separately f) when mice are 6 weeks old they may be mated (see 1.
Alternatively, chimeras can be mated with 129/Sv+Tyr-c+p mice, which have the same genotype as the embryonic stem cells.
This will produce mice with the targeted gene mutation on the 129/Sv+Tyr-c+p background for comparison with the mutation on the C57BL/6 background. Expect litters within a month of mating since female mice go into estrus every 3 or 4 days and the gestation time of mice is 19-21 days.
If you have 10 or so chimeras to take through breeding then these guidelines apply.
If you only have one or two animals then breed them indefinitely until you can produce additional animals from independent ES cell clones.
After 6 generations of mating to C57BL/6, more than 99% of the genetic background will be C57BL/6.By analyzing gene expression on a C57BL/6 background, any influence of the genetic background on gene expression will be controlled for by comparison to normal C57BL/6 mice.