The success of generating new mouse models depends on whether the mES cells in the chimeric organism were also involved in germ cell development. Analysis of sperm from chimeras by short tandem repeat (STR) genotyping can accurately predict the germline inheritance rate after in vitro fertilization (IVF). For this purpose, the proportion of mES cells and the donor mouse strain is determined by genetic fingerprinting.

The samples can be applied to filter paper (e.g. PerkinElmer 226 Spot Saver RUO Card, see figure below) and then dried and labeled accordingly. Shipping can be done at room temperature. Important: In order to precisely locate the invisible sperm adhesion on the filter paper for the subsequent DNA extraction, it is essential to place them in the center of the specified circles! You are on the safe side if the place for the application is marked as a dot with a pencil beforehand. Any remaining circles without sample material should be crossed out.

Preparation of sperm from the epididymis (bilateral): After sperm are collected from the epididymis, they are taken up and mixed in 180 µl of CPA (Cryo Protection Agent). From this, 3-4 µl per filter spot are applied centrally.

In parallel, the STR genotype of a blood sample (without additive, such as EDTA or heparin) from the same animal is used to confirm the genotyping data (sample volume 5 µl - 10 µl per spot). Blood and semen must be submitted in duplicate each.

Note: Not all filter papers are equally suitable for applying the sample material. The PerkinElmer 226 Spot Saver RUO Card shown in the picture has been successfully tested by us. In addition to a favorable price, it also has all the necessary outer packaging for proper sample labeling and contamination-free shipping.

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