Stock Number: 010799 Strain Name: FVB;129S6-Sncatm1Nbm Tg(SNCA*A53T)1Nbm Tg(SNCA*A53T)2Nbm/J

Allele: Tg(SNCA) Protocol Name: Tg(SNCA) Method: Standard PCR Version: 1.0 Created: 13-January -2010 Updated: 03-May -2010

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 9801 - CCT CCT GTT AGC TGG GCT TT - Transgene Forward 9802 - ACC ACT CCC TCC TTG GTT TT - Transgene Reverse oIMR7338 - CTA GGC CAC AGA ATT GAA AGA TCT - Internal Positive Control Forward oIMR7339 - GTA GGT GGA AAT TCT AGC ATC ATC C - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 DNA 2.00 - 2.00 2.5 mM dNTP 0.96 0.20 mM 0.96 ddH2O 3.72 - 3.72 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 20 uM 9801 0.60 1.00 uM 0.60 20 uM 9802 0.60 1.00 uM 0.60 5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06 20 uM oIMR7338 0.60 1.00 uM 0.60 20 uM oIMR7339 0.60 1.00 uM 0.60

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 se - 3 58 30 sec - 4 72 30 sec - 5 - - Go to step 2-4, 35 times 6 72 2 min - 7 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 010710 FVB;129S6-Sncatm1Nbm Tg(SNCA)1Nbm/J 010788 FVB;129S6-Sncatm1Nbm Tg(SNCA*A30P)1Nbm Tg(SNCA*A30P)2Nbm/J 010799 FVB;129S6-Sncatm1Nbm Tg(SNCA*A53T)1Nbm Tg(SNCA*A53T)2Nbm/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.