Stock Number: 008684 Strain Name: B6.Cg-Rag1tm1Mom Tyrp1B-w Tg(Tcra,Tcrb)9Rest/J

Allele: Rag1tm1Mom Protocol Name: Rag1tm1Mom Method: Standard PCR Version: 1.1 Created: 09-February -2001 Updated: 03-March -2010

Notes: An artifact band of ~ 640 bp may be amplified in heterozygous mutant mice. Expected Results: Mutant = 530 bp Heterozygote = 474 bp and 530 bp Wild type = 474 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 oIMR1746 - GAG GTT CCG CTA CGA CTC TG - Wild type Forward oIMR3104 - CCG GAC AAG TTT TTC ATC GT - Common oIMR8162 - TGG ATG TGG AAT GTG TGC GAG - Mutant Forward

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.68 - 3.68 10 X AB PCR BufferII 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 oIMR1746 0.60 1.00 uM 0.60 20 uM oIMR3104 0.30 0.50 uM 0.30 20 uM oIMR8162 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 0.04 0.02 U/ul 0.04 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 2 min - 2 94 30 sec - 3 58 45 sec - 4 72 45 sec repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 002096 B6;129S7-Rag1tm1Mom/J 002216 B6.129S7-Rag1tm1Mom/J 002506 STOCK Tg(CD3E)26Cpt-Rag1tm1Mom/J 002720 AKR/CumJ 003145 C.129S7(B6)-Rag1tm1Mom/J 003729 NOD.129S7(B6)-Rag1tm1Mom/J 004347 NOD.Cg-Rag1tm1Mom Tg(TcraAI4)1Dvs/DvsJ 004369 B6.Cg-Rag1tm1Mom Ins2Akita/J 004673 NOD.129(B6)-Rag1tm1Mom Cd80tm1Shr/JbsJ 004848 NOD.Cg-Rag1tm1Mom Prf1tm1Sdz/SzJ 005707 STOCK Rag1tm1Mom Tg(TIE2-lacZ)182Sato/J 006022 NOD.Cg-Rag1tm1Mom H2-Ab1tm1Gru Tg(CD2-CD4,HLA-DQA1,HLA-DQB1)1Ell/EllJ 006024 NOD.Cg-Rag1tm1Mom H2-Ab1tm1Gru Tg(CD4,HLA-DQA1,HLA-DQB1)N8Ell/EllJ 006770 STOCK Rag1tm1Mom Tg(TIE2GFP)287Sato/J 007799 NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ 008659 NOD.Cg-Rag1tm1Mom Ins2Akita Prf1tm1Sdz/SzJ 008684 B6.Cg-Rag1tm1Mom Tyrp1B-w Tg(Tcra,Tcrb)9Rest/J 016919 B6.Cg-Rag1tm1Mom Fcgrttm1Dcr Tg(CAG-FCGRT)276Dcr/DcrJ 017700 B6.129-Rag1tm1Mom Fcgrttm1Dcr/DcrJ 017914 NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito/ScasJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.