Stock Number: 004223 Strain Name: CHa.SWV(C3Fe)-Mbpshi/J

Allele: Mbpshi Protocol Name: Mbpshi Method: Standard PCR Version: 1.1 Created: 16-September-2008 Updated: 11-June -2010

Notes: The mutant forward and reverse primers flank the Mbp<shi> deletion. This assay will NOT distinguish heterozygous from homozygous mutant animals. Homozygous mutant mice are identifiable phenotypically. Expected Results: Mutant = 308 bp Internal positive control = 200 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 oIMR0968 - ACC GTC CTG AGA CCA TTG TC - Mutant oIMR0969 - GTG CTT ATC TAG TGT ATG CCT GTG - Mutant oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control Forward oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.40 - 3.40 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 oIMR0968 0.60 1.00 uM 0.60 20 uM oIMR0969 0.60 1.00 uM 0.60 20 ul oIMR8744 0.40 0.67 ul 0.40 20 uM oIMR8745 0.40 0.67 uM 0.40 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 001428 C3Fe.SWV-Mbpshi/J 004223 CHa.SWV(C3Fe)-Mbpshi/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.