Stock Number: 002935 Strain Name: FVB.129S2(B6)-Ccnd1tm1Wbg/J

Allele: Ccnd1tm1Wbg Protocol Name: Ccndtm1Wbg Method: Standard PCR Version: 1.1 Created: 22-January -2001 Updated: 13-April -2010

Expected Results: Mutant = 391 bp Heterozygote = 224 bp and 391 bp Wild type = 224 bp

Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0329 - TAG CAG AGA GCT ACA GAC TTC G - Common oIMR0330 - CTC CGT CTT GAG CAT GGC TC - Wild type oIMR0331 - CTA GTG AGA CGT GCT ACT TC - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.76 - 3.76 10 X AB PCR BufferII 0.96 0.80 X 0.96 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR0329 0.60 1.00 uM 0.60 20 uM oIMR0330 0.45 0.75 uM 0.45 20 uM oIMR0331 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 0.05 0.02 U/ul 0.05 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 64 45 sec - 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 002537 B6;129S2-Ccnd1tm1Wbg/J 002935 FVB.129S2(B6)-Ccnd1tm1Wbg/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.