Difference between revisions of "Genotyping Program"

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{| border="1"
 
{| border="1"
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!| Allele || Program|| Fragment Size(s) || Notes
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!| Allele || Program|| Fragment Size(s) || Strain(s) || Notes
 
|-  
 
|-  
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|TSC-floxed || Inoki PCR || || Two bands if heterozygote
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|[[TSC-floxed Genotyping| TSC-floxed]]|| [[Cre PCR]] || KO- 368bp, WT- 295bp || aTSC, mTSC, Placenta TSC || Two bands if heterozygote
 
|-  
 
|-  
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|Cre || Inoki PCR || || Only present if Transgene Positive (cannot detect homozygosity at transgene)
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|[[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)
 
|-
 
|-
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|Vac14 || Inoki PCR|| || Two bands if heterozygote
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|[[SRY Genotyping| SRY]]|| [[Cre PCR ]] || Between 100-200kDa || any mice too young to sex visually || Only present in males
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|-
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|[[Vac14 Genotyping|Vac14]] || [[Cre PCR]]|| || || Two bands if heterozygote
 
|-  
 
|-  
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|TC10b || TC10b || KO - 500bp, WT-1kb || Two bands if heterozygote
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|[[TC10b Genotyping|TC10b]] || TC10b || KO - 500bp, WT-1kb || || Two bands if heterozygote
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|-
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|[[Ob/ob_Genotyping|ob/ob Genotyping]] || ob/ob|| KO - 123bp, WT-104kb, control 191 bp || || Two reactions, see [[Ob/ob_Genotyping|ob/ob Genotyping]]
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|-
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|[[MTM1 Genotyping|MTM1]] || MTM1 || Mutant-575bp, WT-714bp  || || Separate Reactions for WT and KO
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|-
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|[[CAP Genotyping|CAP]] || CAP ||  || || Two bands if heterozygote, upper band for KO
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|-
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|[[Vinexin Genotyping|Vinexin]] || Vinexin || || ||
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|-
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|[[nr3c1 Genotyping| GR KO]] || nr3c1 || KO- 300bp, WT- 247bp || GR-KO and Adipo-GR-KO || Two bands if heterozygote
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|-
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|[[mc4r-Cone Genotyping| mc4r-Cone]] || mc4r || KO- 540bp, WT- 400bp || mc4r || Two bands if heterozygote (run primers separately)
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|-
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|[[ppp1r3c Genotyping| PTG]] || ppp1r3c (PTG) || KO- 574bp, WT- 678bp || ppp1r3c (PTG) || Two bands if heterozygote (run primers separately)
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|-
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|[[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)
 +
|-
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|[[GDF15 Genotyping| GDF15]] || GDF15 || KO- 200bp, WT- 600bp || GDF15 || Two bands if heterozygote (run primers separately)
 
|}
 
|}
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*Bring up to 500 mL with ddH20
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*Sterile filter buffer and store at 4C
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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]]
 
[[ Category:PCR]]
 
[[ Category:PCR]]
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[[ Category:DNA]]
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[[ Category:Molecular Biology]]

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.

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