Stock Number: 006900 Strain Name: B6.129-St3gal4tm1.1Jxm/J

Allele: St3gal4tm1.1Jxm Protocol Name: St3gal4tm1.1Jxm Method: Standard PCR Version: 1.1 Created: 25-September-2007 Updated: 29-September-2009

Expected Results: Mutant = 450 bp Heterozygote = 450 bp and 290 bp Wild type = 290 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 oIMR6890 - GAC GCC ATC CAC CTA TGA G - - oIMR6891 - GGC TGC TCC CAT TCC ACT - - oIMR6892 - GGC TCT TTG TGG GAC CAT CAG - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 10 X AB PCR BufferII 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 oIMR6890 0.60 1.00 uM 0.60 20 uM oIMR6891 0.60 1.00 uM 0.60 20 uM oIMR6892 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 65 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 006900 B6.129-St3gal4tm1.1Jxm/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.