Stock Number: 002361 Strain Name: B6;129S7-Asgr2tm1Her/J

Allele: Asgr2tm1Her Protocol Name: Asgr2 Method: Standard PCR Version: 1.1 Created: 21-April -2009 Updated: 24-September-2009

Expected Results: Mutant = 280 bp Heterozygote = 116 bp and 280 bp Wildtype = 116 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 oIMR1467 - CCT TTC CAG TGT GGA TTT TCT - - oIMR1468 - GGA CTG TCA AGA CAT CCA GCA - - 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 3.16 - 3.16 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 oIMR1467 0.40 0.67 uM 0.40 20 uM oIMR1468 0.40 0.67 uM 0.40 20 uM oIMR6916 0.60 1.00 uM 0.60 20 uM oIMR6917 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 002361 B6;129S7-Asgr2tm1Her/J 002387 129-Asgr2tm1Her/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.