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. Author manuscript; available in PMC: 2020 May 1.
Published in final edited form as: Biochem Pharmacol. 2019 Jan 30;163:21–31. doi: 10.1016/j.bcp.2019.01.022

Figure 2. Genotyping protocol.

Figure 2.

Mice are genotyped using two sets of primers to identify the 5’ LoxP site upstream of exon 1 and the 3’ LoxP site within intron 1. A third primer set is used to detect presence of the Myh6-Cre transgene. Lane 1: The reverse primer of the 5’ LoxP primer set anneals to the LoxP sequence thereby generating an 820 bp amplicon from the floxed allele or no product with the wild-type allele. Lane 2: The 3’ LoxP primer set flanks the 3’ LoxP site thereby generating a 581 bp amplicon diagnostic of the wild-type allele and a 615 bp amplicon diagnostic of the floxed allele. The presence of both amplicons with the 3’ LoxP primer set indicates heterozygosity. Lane 3: The Myh6-Cre primer set amplifies a 300 bp product from the Myh6-Cre transgene.