Stock Number: 000058 Strain Name: B6(Cg)-Tyrc-2J/J

Allele: Tyrc-2J Protocol Name: NntC57BL/6J Method: Standard PCR Version: 1.1 Created: 05-June -2008 Updated: 24-September-2009

Expected Results: Mutant = 750 bp Heterozygote = ~570 bp and 750 bp Wild type = ~570 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 oIMR7332 - GTA GGG CCA ACT GTT TCT GCA TGA - Common oIMR7333 - GGG CAT AGG AAG CAA ATA CCA AGT TG - Wild type oIMR7334 - GTG GAA TTC CGC TGA GAG AAC TCT T - Mutant

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 oIMR7332 0.60 1.00 uM 0.60 20 uM oIMR7333 0.60 1.00 uM 0.60 20 uM oIMR7334 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 60 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 000058 B6(Cg)-Tyrc-2J/J 004353 C57BL/6-Tg(UBC-GFP)30Scha/J 008398 STOCK Fxntm1Mkn Tg(FXN)YG8Pook/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.