Stock Number: 006390 Strain Name: B6;129-Sncatm1Sud Sncbtm1.1Sud/J

Allele: Sncatm1Sud Protocol Name: Sncatm1Sud Method: Standard PCR Version: 1.1 Created: 26-June -2008 Updated: 24-September-2009

Expected Results: Mutant = 300 bp Heterozygote = 320 bp and 300 bp Wild type=320 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 3.0% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6428 - TGC CCC TGA AAT GCT GCG CC - - 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 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 oIMR7967 0.60 1.00 uM 0.60 20 uM oIMR7968 0.60 1.00 uM 0.60 20 uM oIMR6428 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 70 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 006390 B6;129-Sncatm1Sud Sncbtm1.1Sud/J 008132 STOCK Tg(THY1-Snca)M1mSud/J 008135 B6.Cg-Tg(THY1-SNCA*A53T)M53Sud/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.