Stock Number:013149
Strain Name:B6.129-Dysftm1Kcam/J

Dysftm1Kcam
Dysftm1Kcamalternate1
Separated PCR
1.0
23-November -2011 06-January -2014
Expected Results:

Mutant = 207 bp
Heterozygote = 207 bp and 330 bp
Wild type = 330 bp

Gel Image 
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Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
13563 - AAG CTG GAA ATG ACC TTG GA - Wild type Forward
13564 - CAC TAT CCT CCT GCC TCA GC - Wild type Reverse
oIMR7542 - GGA GCT CAG GTG TCC AGT GT - Mutant Forward
oIMR8444 - GCC TGA AGA ACG AGA TCA GC - Mutant Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration
Total Volume (µl)
20 uM 13564 0.30 0.25 uM 0.30
2.5 U/ul Kapa 2G HS taq polymerase 0.05 0.01 U/ul 0.05
25 mM MgCl2 0.96 2.00 mM 0.96
10 mM dNTPS-kapa 0.24 0.20 mM 0.24
20 uM 13563 0.30 0.25 uM 0.30
5 mM 10x Loading Dye 1.20 0.50 mM 1.20
5 X Kapa 2G HS buffer 2.40 1.00 X 2.40
ddH2O 4.55 - 4.55
DNA 2.00 - 2.00
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
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration
Total Volume (µl)
ddH2O 4.55 - 4.55
5 X Kapa 2G HS buffer 2.40 1.00 X 2.40
25 mM MgCl2 0.96 2.00 mM 0.96
10 mM dNTPS-kapa 0.24 0.20 mM 0.24
20 uM oIMR7542 0.30 0.25 uM 0.30
20 uM oIMR8444 0.30 0.25 uM 0.30
5 mM 10x Loading Dye 1.20 0.50 mM 1.20
2.5 U/ul Kapa 2G HS taq polymerase 0.05 0.01 U/ul 0.05
DNA 2.00 - 2.00
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
006830 129-Dysftm1Kcam/J
013149 B6.129-Dysftm1Kcam/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 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