Stock Number: 007251 Strain Name: B6.129-Mapttm1Hnd/J

Allele: Mapttm1Hnd Protocol Name: Mapttm1Hnd Method: Standard PCR Version: 1.1 Created: 04-July -2008 Updated: 24-September-2009

Expected Results: Mutant = ~170 bp Heterozygote = ~170 bp and 269 bp Wild type = 269 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 oIMR7415 - GCC AGA GGC CAC TTG TGT AG - Mutant oIMR7824 - AAT GGA AGA CCA TGC TGG AG - Wild type Forward oIMR7825 - ATT CAA CCC CCT CGA ATT TT - Common

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 oIMR7824 0.60 1.00 uM 0.60 20 uM oIMR7825 0.60 1.00 uM 0.60 20 uM oIMR7415 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 57 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 007251 B6.129-Mapttm1Hnd/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.