Genotyping Program

From Bridges Lab Protocols
Jump to: navigation, search
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.