Stock Number: 006953 Strain Name: STOCK Notch1tm3(cre)Rko/J

Allele: Notch1tm3(cre)Rko Protocol Name: Notch1tm3(cre)Rko Method: Standard PCR Version: 1.1 Created: 13-April -2009 Updated: 24-September-2009

Expected Results: HET = 181 bp and 450 bp MUT = 450 bp WT = 181 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 oIMR9375 - TCG TGC TCC TGT TCT TTG TG - Common oIMR9376 - CAC ACA GTG GGA GCA GTG TC - Wild type Reverse oIMR9377 - ATG TTT AGC TGG CCC AAA TG - Mutant Reverse

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 oIMR9375 0.60 1.00 uM 0.60 20 uM oIMR9376 0.60 1.00 uM 0.60 20 uM oIMR9377 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 64 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 006953 STOCK Notch1tm3(cre)Rko/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.