Stock Number: 002445 Strain Name: STOCK Notch1tm1Con/J

Allele: Notch1tm1Con Protocol Name: Notch1tm1Con Method: Standard PCR Version: 1.1 Created: 16-June -2008 Updated: 24-September-2009

Expected Results: Mutant = ~400 bp Heterozygote = ~300 bp and ~400 bp Wild type = ~300 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 oIMR0275 - GAT ATC GTG GTG CAT ACC CTC CTG - Common oIMR0276 - GTG GTC TAG GAT GCT TGG GTC TAG - Wild type oIMR6489 - GTC AGT TTC ATA GCC TGA AGA ACG - 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 oIMR0275 0.60 1.00 uM 0.60 20 uM oIMR0276 0.60 1.00 uM 0.60 20 uM oIMR6489 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 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 002445 STOCK Notch1tm1Con/J 002797 B6.129-Notch1tm1Con/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.