Genotyping Program

Allele	Program	Fragment Size(s)	Strain(s)	Notes
TSC-floxed	Cre PCR	KO- 368bp, WT- 295bp	aTSC, mTSC, Placenta TSC	Two bands if heterozygote
Cre	Cre PCR	Between 340-400kDa	aTSC, mTSC, Adipo-Raptor, Adipo-GR-KO, Placenta TSC, Placenta GR	Only present if Transgene Positive (cannot detect homozygosity at transgene)
SRY	Cre PCR	Between 100-200kDa	any mice too young to sex visually	Only present in males
Vac14	Cre PCR			Two bands if heterozygote
TC10b	TC10b	KO - 500bp, WT-1kb		Two bands if heterozygote
ob/ob Genotyping	ob/ob	KO - 123bp, WT-104kb, control 191 bp		Two reactions, see ob/ob Genotyping
MTM1	MTM1	Mutant-575bp, WT-714bp		Separate Reactions for WT and KO
CAP	CAP			Two bands if heterozygote, upper band for KO
Vinexin	Vinexin
GR KO	nr3c1	KO- 300bp, WT- 247bp	GR-KO and Adipo-GR-KO	Two bands if heterozygote
mc4r-Cone	mc4r	KO- 540bp, WT- 400bp	mc4r	Two bands if heterozygote (run primers separately)
PTG	ppp1r3c (PTG)	KO- 574bp, WT- 678bp	ppp1r3c (PTG)	Two bands if heterozygote (run primers separately)
4EBP1/2	Cre PCR	1= KO- 550bp, WT- 175bp; 2= KO- 750bp, WT- 450bp	4EBP1/2	Two bands if heterozygote for each allele (run primers for 1 and 2 separately)
GDF15	GDF15	KO- 200bp, WT- 600bp	GDF15	Two bands if heterozygote (run primers separately)

AMPK, fTSC, fRaptor, and C57Bl6/J are all wild-type mice and don't need to be genotyped.