Stock Number:008374
Strain Name:C57BL/6-Foxp3tm1Flv/J
Foxp3tm1Flv
Foxp3tm1Flv
Separated PCR
1.1
23-September-2008 24-September-2009
Notes: The Foxp3 gene is X-LINKED. Males with the targeted allele are hemizygous mutants
Expected Results: Mutant = 470 bp
Heterozygote = 470 bp and 510 bp
Wild type = 510 bp
Gel Image 
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Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
oIMR8611 - CAA AAC CAA GAA AAG GTG GGC - Common
oIMR8612 - CAG TGC TGT TGC TGT GTA AGG GTC - Wild type Reverse
oIMR8613 - GGA ATG CTC GTC AAG AAG ACA GG - Mutant Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 3.96 - 3.96
10 X AB PCR BufferII 1.20 1.00 X 1.20
25 mM MgCl2 0.96 2.00 mM 0.96
2.5 mM dNTP 0.96 0.20 mM 0.96
20 uM oIMR8611 0.60 1.00 uM 0.60
20 uM oIMR8613 0.60 1.00 uM 0.60
5 mM DNA Loading Dye 1.66 0.69 mM 1.66
5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06
DNA 2.00 - 2.00
Cycling 
Step # Temp °C Time Note
1 94 3 min -
2 94 30 sec -
3 64 1 min -
4 72 1 min repeat steps 2-4 for 35 cycles
5 72 2 min -
6 10 - hold
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 3.96 - 3.96
10 X AB PCR BufferII 1.20 1.00 X 1.20
25 mM MgCl2 0.96 2.00 mM 0.96
2.5 mM dNTP 0.96 0.20 mM 0.96
20 uM oIMR8611 0.60 1.00 uM 0.60
20 uM oIMR8612 0.60 1.00 uM 0.60
5 mM DNA Loading Dye 1.66 0.69 mM 1.66
5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06
DNA 2.00 - 2.00
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
008374 C57BL/6-Foxp3tm1Flv/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.
Cycling 
Step # Temp °C Time Note
1 94 3 min -
2 94 30 sec -
3 64 1 min -
4 72 1 min repeat steps 2-4 for 35 cycles
5 72 2 min -
6 10 - hold