Stock Number: 004919 Strain Name: B6.Cg-Fcgrttm1Dcr Tg(CAG-FCGRT)276Dcr/DcrJ

Allele: Fcgrttm1Dcr Protocol Name: Fcgrttm1Dcr Method: Standard PCR Version: 1.1 Created: 16-June -2008 Updated: 21-October -2009

Expected Results: Mutant = 378 bp Heterozygote = 248 bp and 378 bp Wild type = 248 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 oIMR7720 - GGG ATG CCA CTG CCC TG - Wild type Forward oIMR7721 - CGA GCC TGA GAT TGT CAA GTG TAT T - Common oIMR7722 - GGA ATT CCC AGT GAA GGG C - Mutant Forward

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 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 oIMR7720 0.60 1.00 uM 0.60 20 uM oIMR7721 0.60 1.00 uM 0.60 20 uM oIMR7722 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 59 1 min - 4 72 1 min 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 003982 B6.129X1-Fcgrttm1Dcr/DcrJ 003986 MRL.129X1-Fcgrttm1Dcr/DcrJ 003988 NOD.129X1-Fcgrttm1Dcr/DvsJ 003989 NZM2410.129X1-Fcgrttm1Dcr/DcrJ 003990 SJL.129X1-Fcgrttm1Dcr/DcrJ 004919 B6.Cg-Fcgrttm1Dcr Tg(CAG-FCGRT)276Dcr/DcrJ 014565 B6.Cg-Fcgrttm1Dcr Tg(FCGRT)32Dcr/DcrJ 016919 B6.Cg-Rag1tm1Mom Fcgrttm1Dcr Tg(CAG-FCGRT)276Dcr/DcrJ 017700 B6.129-Rag1tm1Mom Fcgrttm1Dcr/DcrJ 018441 B6.Cg-Fcgrttm1Dcr Prkdcscid Tg(FCGRT)32Dcr/DcrJ 018541 B6.Cg-Fcgrttm1Dcr Prkdcscid/DcrJ 021146 B6.Cg-Fcgrttm1Dcr Prkdcscid Tg(CAG-FCGRT)276Dcr/DcrJ 374176 B6.129X1-Fcgrttm1Dcr/DcrJ X B6.Cg-Fcgrttm1DcrTg(FCGRT)276Dcr Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.