Stock Number: 008133 Strain Name: B6.129-Sncbtm1Sud/J

Allele: Sncbtm1Sud Protocol Name: Sncbtm1Sud Method: Standard PCR Version: 1.1 Created: 08-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 360 bp Heterozygote = 316 bp and 360 bp Wild type = 316 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 oIMR7967 - AGG ACA CCA CTG GCC CCG AGT CC - Forward oIMR7968 - GAC GCA CGT CCG CAC GTC CAC CC - Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.86 - 4.86 10 X PCR Buffer II (ABI) 0.96 0.80 X 0.96 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR7967 0.60 1.00 uM 0.60 20 uM oIMR7968 0.60 1.00 uM 0.60 5 M Betaine 1.00 0.42 M 1.00 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 68 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 008133 B6.129-Sncbtm1Sud/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.