Stock Number: 003615 Strain Name: B6.129-Psen1tm1Shn/J

Allele: Psen1tm1Shn Protocol Name: Psen1tm1Shn Method: Standard PCR Version: 1.1 Created: 28-August -2008 Updated: 24-September-2009

Expected Results: Mutant = 300 bp Heterozygote = 300 bp and 345 bp Wild type = 345 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 3.0% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1245 - ACC TCA GCT GTT TGT CCC GG - Wild type oIMR1247 - TCT GGA AGT AGG ACA AAG GTG - Wild type oIMR7696 - GCA CGA GAC TAG TGA GAC GTG - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.38 - 3.38 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 oIMR1245 0.60 1.00 uM 0.60 20 uM oIMR7696 0.60 1.00 uM 0.60 20 uM oIMR1247 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 0.04 0.02 U/ul 0.04 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 65 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 003615 B6.129-Psen1tm1Shn/J 003822 B6;129S-Psen1tm1Shn/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.