Stock Number: 008159 Strain Name: STOCK Gt(ROSA)26Sortm1(Notch1)Dam/J

Allele: Gt(ROSA)26Sortm1(Notch1)Dam Protocol Name: Gt(ROSA)26Sortm1(Notch1)Dam Method: Standard PCR Version: 1.1 Created: 11-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 320 bp Heterozygote = 235 bp and 320 bp Wild type = 235 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 oIMR8038 - TAA GCC TGC CCA GAA GAC TC - Wild type Reverse oIMR8039 - GAA AGA CCG CGA AGA GTT TG - Mutant Reverse oIMR8545 - AAA GTC GCT CTG AGT TGT TAT - 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 oIMR8545 0.60 1.00 uM 0.60 20 uM oIMR8038 0.60 1.00 uM 0.60 20 uM oIMR8039 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 54 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 008159 STOCK Gt(ROSA)26Sortm1(Notch1)Dam/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.