Transgenic animals are produced by injecting a DNA construct directly into the pronucleus of a newly fertilized mouse egg. Plasmid or BAC constructs can be used. We routinely inject eggs from C57Bl6 inbred mice. In some eggs, the DNA will integrate into the mouse chromosomal DNA before cell division, and will be present in all cells in the mouse including the germ line. The Transgenic Animal Production service consists of injecting each construct into eggs from fifteen females per day. We suggest starting with three sessions. Fifteen to twenty mice will normally be born from the three sessions. These animals are screened for the presence of the transgene by a polymerase chain reaction genotyping assay.
The number of transgenic animals typically varies from two to four from three sessions. When the founder animals are at weaning age, they are transferred to the investigator. For all constructs, the investigator must provide the transgenic construct prepared by a suitable protocol, as well as a set of oligonucleotides primers suitable for PCR genotyping by our Universal Genotyping Assay. Primers for this assay must be 30 nucleotides in length, have 40-60% GC content, have non-homologous 3’ ends, contain no repetitive sequences, and produce an amplimer of 100-500 base pairs in length. We are happy to help with primer design if you provide the transgene sequence, or you can see the Universal Genotyping Assay for more information.
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