Stock Number: 008135 Strain Name: B6.Cg-Tg(THY1-SNCA*A53T)M53Sud/J

Allele: Tg(THY1-SNCA*A53T)M53Sud Protocol Name: Tg(THY1-SNCA*A53T)M53Sud, F53Sud Method: Standard PCR Version: 2.0 Created: 26-May -2009 Updated: 24-September-2009

Expected Results: Transgene = 500 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 oIMR8584 - GGC ACC TAG AGG ATC TCG ACT AGT GG - Forward oIMR8585 - GGA CCT CGA CGC TTA GGC TTC AGG - Reverse oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control Forward oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.06 - 4.06 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 oIMR8584 0.60 1.00 uM 0.60 20 uM oIMR8585 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 20 oIMR8744 0.40 0.67 0.40 20 uM oIMR8745 0.40 0.67 uM 0.40

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 30 sec - 3 62 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 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.