Stock Number: 007708 Strain Name: B6.129-Gt(ROSA)26Sortm1(HD*103Q)Xwy/J

Allele: Gt(ROSA)26Sortm1(HD*103Q)Xwy Protocol Name: Gt(ROSA)26Sortm1(HD*103Q)Xwy Method: Standard PCR Version: 1.1 Created: 12-August -2008 Updated: 24-September-2009

Expected Results: Mutant = ~300 bp Heterozygote = ~300 bp and 603 bp Wild type = 603 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 oIMR8052 - GCG AAG AGT TTG TCC TCA ACC - Mutant oIMR8545 - AAA GTC GCT CTG AGT TGT TAT - Common oIMR8546 - GGA GCG GGA GAA ATG GAT ATG - Wild type Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.60 - 3.60 10 X PCR Buffer II (ABI) 0.96 0.80 X 0.96 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR8546 0.60 1.00 uM 0.60 20 uM oIMR8545 0.60 1.00 uM 0.60 20 uM oIMR8052 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 5 min - 2 94 30 sec - 3 60 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 007708 B6.129-Gt(ROSA)26Sortm1(HD*103Q)Xwy/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.