Stock Number: 007453 Strain Name: B6.129P2(Cg)-Dhcr7tm1Gst/J

Allele: Dhcr7tm1Gst Protocol Name: Dhcr7tm1Gst Method: Standard PCR Version: 1.1 Created: 07-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 370 bp Heterozygote = 370 bp and 425 bp Wild type = 425 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 oIMR7011 - CCG CCA CTT CAA CTA TAC TGG TGA C - - oIMR7013 - CAT TGC TCA GCG GTG CTG TC - - oIMR7014 - GCC ATT CTG CTG GGG TAT CC - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 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 oIMR7011 0.60 1.00 uM 0.60 20 uM oIMR7013 0.60 1.00 uM 0.60 20 uM oIMR7014 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 60 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 007453 B6.129P2(Cg)-Dhcr7tm1Gst/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.