Stock Number: 008364 Strain Name: B6;129-Chattm1(cre/Esr1)Nat/J

Allele: Chattm1(cre/Esr1)Nat Protocol Name: Chattm1(cre/Esr1) Method: Standard PCR Version: 1.1 Created: 15-September-2008 Updated: 24-September-2009

Expected Results: Mutant = 630 bp Heterozygote = 540 bp and 630 bp Wild type = 540 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 oIMR8398 - AAG AGC TAG AGG CCC AAC CAA GCC A - Wild type Forward oIMR8399 - TTC TGA ATA GCT GGA TGA GAT AAT C - Common oIMR8703 - CGC ATA GAA ATT GCA TCA ACG CAT - Mutant Forward

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 oIMR8398 0.60 1.00 uM 0.60 20 uM oIMR8399 0.60 1.00 uM 0.60 20 uM oIMR8703 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 61.5 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 008364 B6;129-Chattm1(cre/Esr1)Nat/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.