Stock Number: 008473 Strain Name: B6;SJL-Tg(THY1-SNCA*A30P)M30Sud/J

Allele: Tg(THY1-SNCA*A30P)M30Sud Protocol Name: Tg(THY1-SNCA*A30P)M30Sud Method: Standard PCR Version: 1.1 Created: 25-August -2008 Updated: 24-September-2009

Expected Results: TG = 500 bp internal control = 200 bp

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 4.26 - 4.26 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 20 oIMR8744 0.30 0.50 0.30 20 uM oIMR8745 0.30 0.50 uM 0.30 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 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 008473 B6;SJL-Tg(THY1-SNCA*A30P)M30Sud/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.