Stock Number: 002463 Strain Name: B6.129S-Itga4tm1Hyn/J

Allele: Itga4tm1Hyn Protocol Name: Itga4tm1Hyn Method: Standard PCR Version: 1.1 Created: 21-May -2003 Updated: 24-September-2009

Expected Results: Mutant = 280 bp Heterozygote = 324 bp and 280 bp Wild type = 324 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 oIMR0563 - AGC CTG AAG CAT CCC TGG - Wild type oIMR0564 - GGT CCA GGT TGT AGG AGT G - Wild type oIMR6916 - CTT GGG TGG AGA GGC TAT TC - Mutant Forward oIMR6917 - AGG TGA GAT GAC AGG AGA TC - Mutant 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 oIMR0563 0.60 1.00 uM 0.60 20 uM oIMR0564 0.60 1.00 uM 0.60 20 uM oIMR6916 0.30 0.50 uM 0.30 20 uM oIMR6917 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 60 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 002463 B6.129S-Itga4tm1Hyn/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.