Stock Number:006941
Strain Name:B6.129S7-B4galt1tm1Shur/J
B4galt1tm1Shur
B4galt1tm1Shur
Separated PCR
1.1
09-July -2008 24-September-2009
Expected Results: Mutant = ~242 bp
Heterozygote = ~242 bp and 640 bp
Wild type = 640 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
oIMR8115 - AGA CTC CGC CCC CCC AGG TT - Wild type Forward
oIMR8116 - CTC ACG AAA CCT CAT CTT CC - Wild type Reverse
oIMR8118 - TGG ATG TGG AAT GTG TGC GA - Mutant Reverse
oIMR8203 - AGA AGC TGC ACC AGG GCT GGA - Mutant Forward
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 3.96 - 3.96
10 X PCR Buffer II (ABI) 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 oIMR8203 0.60 1.00 uM 0.60
20 uM oIMR8118 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 56 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 PCR Buffer II (ABI) 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 oIMR8115 0.60 1.00 uM 0.60
20 uM oIMR8116 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
006941 B6.129S7-B4galt1tm1Shur/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 56 1 min -
4 72 1 min repeat steps 2-4 for 35 cycles
5 72 2 min -
6 10 - hold