Stock Number: 006201 Strain Name: B6.129-Scd1tm1Ntam/J

Allele: Scd1tm1Ntam Protocol Name: Scd1tm1Ntam Method: Standard PCR Version: 1.1 Created: 24-June -2008 Updated: 24-September-2009

Expected Results: Mutant = 425 bp Heterozygote = 425 bp and 600 bp Wild type = 600 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 oIMR5361 - GGG TGA GCA TGG TGC TCA GTC CCT - - oIMR5362 - CAC ACC ATA TCT GTC CCC GAC AAA TGT C - - oIMR6982 - ATA GCA GGC ATG CTG GGG AT - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.66 - 3.66 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 oIMR5361 0.60 1.00 uM 0.60 20 uM oIMR5362 0.60 1.00 uM 0.60 20 uM oIMR6982 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 62 30 sec - 4 72 30 sec 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 006201 B6.129-Scd1tm1Ntam/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.