Stock Number: 008843 Strain Name: B6.129P2-Sncgtm1Vlb/J

Allele: Sncgtm1Vlb Protocol Name: Sncgtm1Vlb Method: Standard PCR Version: 1.1 Created: 06-April -2009 Updated: 24-September-2009

Expected Results: HET = 499 bp and 380 bp MUT = 380 bp WT = 499 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 oIMR7412 - ATT AAG GGC CAG CTC ATT CC - Mutant oIMR9583 - AAT CCT GGC CTG GTA CTC TG - Common oIMR9584 - CGG TAC CAA GTT CTC CCA TC - Wild type 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 oIMR9583 0.60 1.00 uM 0.60 20 uM oIMR9584 0.60 1.00 uM 0.60 20 uM oIMR7412 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 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 008843 B6.129P2-Sncgtm1Vlb/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.