Stock Number:006944
Strain Name:B6.129-Mgl1tm1Hed/J
Mgl1tm1Hed
Mgl1tm1Hed
Separated PCR
1.1
10-July -2008 24-September-2009
Notes: This assay does not work well without the use of a Hotstart (We are using Taq Start Antibody mixed 1:1 with Taq polymerase).
Expected Results: Mutant = ~650 bp
Heterozygote = ~650 bp and 1400 bp
Wild type = 1400 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
oIMR8550 - GCG GTT ATT GGA TCT CCT GG - Common
oIMR8551 - CTC AGG TTC CCT CTG TGC CAG C - Wild type Reverse
oIMR8552 - GAC GAG TTC TTC TGA GGG G - Mutant Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 3.78 - 3.78
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 oIMR8550 0.60 1.00 uM 0.60
20 uM oIMR8552 0.30 0.50 uM 0.30
5 M Betaine 2.00 0.83 M 2.00
5 U/ul Taq DNA Polymerase + TaqStart AB (1:1) 0.20 0.08 U/ul 0.20
DNA 2.00 - 2.00
Cycling 
Step # Temp °C Time Note
1 94 3 min -
2 94 30 sec -
3 51 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 3.78 - 3.78
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 oIMR8550 0.60 1.00 uM 0.60
20 uM oIMR8551 0.30 0.50 uM 0.30
5 M Betaine 2.00 0.83 M 2.00
5 U/ul Taq DNA Polymerase + TaqStart AB (1:1) 0.20 0.08 U/ul 0.20
DNA 2.00 - 2.00
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
006944 B6.129-Mgl1tm1Hed/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 66 1 min -
4 72 1 min repeat steps 2-4 for 35 cycles
5 72 2 min -
6 10 - hold**