Difference between revisions of "Genotyping Program"
From Bridges Lab Protocols
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| − | !| Allele || Program|| Fragment Size(s) || Notes | + | !| Allele || Program|| Fragment Size(s) || Strain(s) || Notes |
|- | |- | ||
| − | |[[TSC-floxed Genotyping| TSC-floxed]]|| [[Cre PCR]] || KO- 368bp, WT- 295bp || 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 ]] || || 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) |
|- | |- | ||
| − | |[[Vac14 Genotyping|Vac14]] || [[Cre PCR]]|| | + | |[[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 | ||
|- | |- | ||
| − | |[[TC10b Genotyping|TC10b]] || TC10b || KO - 500bp, WT-1kb || Two bands if heterozygote | + | |[[TC10b Genotyping|TC10b]] || TC10b || KO - 500bp, WT-1kb || || Two bands if heterozygote |
|- | |- | ||
| − | |[[Ob/ob_Genotyping|ob/ob Genotyping]] || ob/ob|| KO - 123bp, WT-104kb, control 191 bp || Two reactions, see [[Ob/ob_Genotyping|ob/ob Genotyping]] | + | |[[Ob/ob_Genotyping|ob/ob Genotyping]] || ob/ob|| KO - 123bp, WT-104kb, control 191 bp || || Two reactions, see [[Ob/ob_Genotyping|ob/ob Genotyping]] |
|- | |- | ||
| − | |[[MTM1 Genotyping|MTM1]] || MTM1 || Mutant-575bp, WT-714bp || Separate Reactions for WT and KO | + | |[[MTM1 Genotyping|MTM1]] || MTM1 || Mutant-575bp, WT-714bp || || Separate Reactions for WT and KO |
|- | |- | ||
| − | |[[CAP Genotyping|CAP]] || CAP || || Two bands if heterozygote, upper band for KO | + | |[[CAP Genotyping|CAP]] || CAP || || || Two bands if heterozygote, upper band for KO |
|- | |- | ||
| − | |[[Vinexin Genotyping|Vinexin]] || Vinexin || || | + | |[[Vinexin Genotyping|Vinexin]] || Vinexin || || || |
|- | |- | ||
| − | |[[nr3c1 Genotyping| GR KO]] || nr3c1 || KO- 300bp, WT- 247bp || Two bands if heterozygote | + | |[[nr3c1 Genotyping| GR KO]] || nr3c1 || KO- 300bp, WT- 247bp || GR-KO and Adipo-GR-KO || Two bands if heterozygote |
|- | |- | ||
| − | |[[mc4r-Cone Genotyping| mc4r-Cone]] || mc4r || KO- 540bp, WT- 400bp || Two bands if heterozygote (run primers separately) | + | |[[mc4r-Cone Genotyping| mc4r-Cone]] || mc4r || KO- 540bp, WT- 400bp || mc4r || Two bands if heterozygote (run primers separately) |
|- | |- | ||
| − | |[[ppp1r3c Genotyping| PTG]] || ppp1r3c (PTG) || KO- 574bp, WT- 678bp || Two bands if heterozygote (run primers separately) | + | |[[ppp1r3c Genotyping| PTG]] || ppp1r3c (PTG) || KO- 574bp, WT- 678bp || ppp1r3c (PTG) || Two bands if heterozygote (run primers separately) |
|- | |- | ||
| − | |[[4EBP1/2 Genotyping| 4EBP1/2]] || [[Cre PCR]] || 1= KO- 550bp, WT- 175bp; 2= KO- 750bp, WT- 450bp || Two bands if heterozygote for each allele (run primers for 1 and 2 separately) | + | |[[4EBP1/2 Genotyping| 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 Genotyping| GDF15]] || GDF15 || KO- 200bp, WT- 600bp || Two bands if heterozygote (run primers separately) | + | |[[GDF15 Genotyping| 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. | |
[[ Category:Genotyping ]] | [[ Category:Genotyping ]] | ||
[[ Category:Mouse Work]] | [[ Category:Mouse Work]] | ||
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.
