Stock Number: 010719 Strain Name: B6;129P2-Prdm9tm1Ymat/J

Allele: Prdm9tm1Ymat Protocol Name: Prdm9tm1Ymat Method: Standard PCR Version: 1.0 Created: 02-August -2009 Updated: 06-July -2010

Expected Results: Mutant = ~470bp Heterozygote = ~470 bp and 319 bp Wild type = 319 bp

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 9118 - AGG AAT CTT CCT TCC TTG CTG TCG - Common 9119 - ATT TCC CTG TAT CTT CTT CAG GAC T - Wild type Reverse 9120 - CGC CAT TCA GGC TGC GCA ACT GTT - Mutant Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06 2.5 mM dNTP 0.96 0.20 mM 0.96 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 25 mM MgCl2 0.96 2.00 mM 0.96 DNA 2.00 - 2.00 20 uM 9119 0.60 1.00 uM 0.60 20 uM 9118 0.60 1.00 uM 0.60 20 uM 9120 0.60 1.00 uM 0.60 10 X AB PCR BufferII 1.20 1.00 X 1.20

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 62 30 sec - 4 72 30 sec - 5 - - Go to step 2-4, 35 times 6 72 2 min - 7 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 010719 B6;129P2-Prdm9tm1Ymat/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.