Stock Number: 008132 Strain Name: STOCK Tg(THY1-Snca)M1mSud/J

Allele: (THY1-Snca)M1mSud Protocol Name: Tg(THY1-Snca) Method: Standard PCR Version: 1.1 Created: 11-July -2008 Updated: 24-September-2009

Expected Results: TG = 490 bp internal control = 200 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 oIMR8420 - GCA TGC TCG AGT TAG GCT TCA GG - Reverse oIMR8584 - GGC ACC TAG AGG ATC TCG ACT AGT GG - Forward 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 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 oIMR8584 0.60 1.00 uM 0.60 20 uM oIMR8420 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 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 64 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 008132 STOCK Tg(THY1-Snca)M1mSud/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.