Stock Number: 008710 Strain Name: B6.129P2(129S4)-Hprttm10(Ple162-EGFP/cre)Ems/Mmjax

Allele: Hprttm10(Ple162-EGFP/cre)Ems Protocol Name: Hprt1tm10(Ple162-EGFP/cre)Ems STD PCR Method: Standard PCR Version: 2.0 Created: 03-November -2009 Updated: 03-November -2009

Expected Results: wt = 370bp het = 370bp & 401bp mut = 401bp

Gel Image

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Gel Information: Use 3% gel

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR9426 - TGT TCT CCT CCA ACC AAA AGG C - - oIMR9427 - TAA AAG GGA TCG GCT GGC AG - - oIMR9435 - AGA TGA GCT TGG TTG CTG GAG TG - - oIMR9436 - TGC TGG ACA TCC CTA CTA ACC CA - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 25 mM MgCl2 0.96 2.00 mM 0.96 ddH2O 2.74 - 2.74 20 uM oIMR9427 0.60 1.00 uM 0.60 DNA 2.00 - 2.00 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR9426 0.60 1.00 uM 0.60 20 uM oIMR9436 0.60 1.00 uM 0.60 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 10 X Platinum Buffer 1.20 1.00 X 1.20 5 U/ul High Fidelity Platinum Taq 0.08 0.03 U/ul 0.08 20 uM oIMR9435 0.60 1.00 uM 0.60

Cycling  Step # Temp °C Time Note 1 94 3min - 2 94 30sec - 3 69 1min - 4 72 1min repeat steps 2-4 for 35 cycles 5 72 2min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 008710 B6.129P2(129S4)-Hprttm10(Ple162-EGFP/cre)Ems/Mmjax Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.