Stock Number:008525
Strain Name:B6.Cg-Tg(Cyp21a1-Apoe)619Fet/J
Tg(Cyp21a1-Apoe)619Fet
Tg(Cyp21a1-Apoe)
Separated PCR
1.1
02-April -2009 24-September-2009
Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals.
Expected Results: Mutant = 733 bp
Heterozygote = 547 bp and 733 bp
Wild type = 547 bp
Gel Image 
Image Image file
View Full Size
Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
oIMR8935 - ACC TCT TTC CTC TGC CCT GCT G - Wild type Forward
oIMR8936 - GCC ATG AGG AGC CAC AGT TTG AAA C - Common
oIMR8937 - CTC CAG TGG ACT CTC CGC CTA CTT G - Transgene Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 4.82 - 4.82
10 X High Fidelity Buffer 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 oIMR8935 0.60 1.00 uM 0.60
20 uM oIMR8936 0.60 1.00 uM 0.60
DMSO 0.80 - 0.80
5 U/ul High Fidelity Platinum Taq 0.06 0.03 U/ul 0.06
DNA 2.00 - 2.00
Cycling 
Step # Temp °C Time Note
1 94 1.5 min -
2 94 30 sec -
3 62 45 sec -
4 72 45 sec 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 5.12 - 5.12
10 X High Fidelity Buffer 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 oIMR8936 0.60 1.00 uM 0.60
20 uM oIMR8937 0.30 0.50 uM 0.30
DMSO 0.80 - 0.80
5 U/ul High Fidelity Platinum Taq 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
008525 B6.Cg-Tg(Cyp21a1-Apoe)619Fet/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 1.5 min -
2 94 30 sec -
3 62 45 sec -
4 72 45 sec repeat steps 2-4 for 35 cycles
5 72 2 min -
6 10 - hold