Difference between revisions of "Genotyping Program"
From Bridges Lab Protocols
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|[[TSC-floxed Genotyping| TSC-floxed]]|| [[Cre PCR]] || KO- 368bp, WT- 295bp || aTSC, mTSC, Placenta TSC || Two bands if heterozygote | |[[TSC-floxed Genotyping| TSC-floxed]]|| [[Cre PCR]] || KO- 368bp, WT- 295bp || aTSC, mTSC, Placenta TSC || Two bands if heterozygote | ||
|- | |- | ||
| − | |[[Cre Genotyping|Cre]] || [[Cre PCR ]] || || aTSC, mTSC, Adipo-Raptor, Adipo-GR-KO, Placenta TSC, Placenta GR || Only present if Transgene Positive (cannot detect homozygosity at transgene) | + | |[[Cre Genotyping|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 Genotyping| SRY]]|| [[Cre PCR ]] || Between 100-200kDa || any mice too young to sex visually || Only present in males | ||
|- | |- | ||
|[[Vac14 Genotyping|Vac14]] || [[Cre PCR]]|| || || Two bands if heterozygote | |[[Vac14 Genotyping|Vac14]] || [[Cre PCR]]|| || || Two bands if heterozygote | ||
Latest revision as of 14:32, 23 July 2021
| 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.
