Stock Number:017762
Strain Name:B6(Cg)-Cacna1ftm1.2Sdie/J
Cacna1ftm1.2Sdie
Cacna1ftm1.2Sdie
Standard PCR
1.0
21-February -2012 21-March -2012
Expected Results:

Mutant = 449 bp
Heterozygote = 290 bp and 449 bp
Wild type = 290 bp

X-linked mutation

 
Gel Image 
Image Image file
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Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
14151 - CAC TCC AGA CAT CCT GCT GA - Forward
14152 - GTC AAC CCA TGC TGT CTC CT - Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
5 X Kapa 2G HS buffer 2.40 1.00 X 2.40
ddH2O 4.55 - 4.55
5 mM 10x Loading Dye 1.20 0.50 mM 1.20
DNA 2.00 - 2.00
10 mM dNTPS-kapa 0.24 0.20 mM 0.24
25 mM MgCl2 0.96 2.00 mM 0.96
2.5 ul Kapa 2G HS taq polymerase 0.05 0.01 ul 0.05
20 uM 14152 0.30 0.50 uM 0.30
20 uM 14151 0.30 0.50 uM 0.30
Cycling 
Step # Temp °C Time Note
1 94 2 min -
2 94 20sec -
3 65 15sec -1.5 C per cycle
4 68 10sec -
5 - - Go to step 2-4, 10 times
6 94 15sec -
7 50 15sec -
8 72 10sec -
9 - - Go to step 6-8, 28 times
10 72 2 min -
11 10 - hold
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
017762 B6(Cg)-Cacna1ftm1.2Sdie/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.