Stock Number: 010818 Strain Name: B6.129-Ifnb1tm1Lky/J

Allele: Ifnb1tm1(EYFP)Lky Protocol Name: Ifnb1tm1(EYFP)Lky Method: Standard PCR Version: 1.0 Created: 21-August -2009 Updated: 12-August -2010

Expected Results: Mutant = 507 bp Heterozygote = 507 bp and 818 bp Wild type = 818 bp

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 9247 - CCC TAT GGA GAT GAC GGA GAA GAT GC - Wild type Forward 9248 - CAA TCC CAT AGC AGG TGA TGC C - Common 9249 - TGG TCC TGC TGG AGT TCG TGA CCG C - Mutant Forward

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.30 - 3.30 5 U/ul Taq DNA Polymerase 0.12 0.05 U/ul 0.12 2.5 mM dNTP 0.96 0.20 mM 0.96 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 25 mM MgCl2 0.96 2.00 mM 0.96 DNA 2.00 - 2.00 20 uM 9247 0.60 1.00 uM 0.60 20 uM 9248 0.60 1.00 uM 0.60 20 uM 9249 0.60 1.00 uM 0.60 10 X AB PCR BufferII 1.20 1.00 X 1.20

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 66 1 min - 4 72 1 min - 5 - - Go to step 2-4, 35 times 6 72 2 min - 7 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 010818 B6.129-Ifnb1tm1Lky/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.