Stock Number: 009043 Strain Name: 129S-Gt(ROSA)26Sortm2Tyj/J

Allele: Gt(ROSA)26Sortm2Tyj Protocol Name: Gt(ROSA)26Sortm2Tyj Method: Standard PCR Version: 1.1 Created: 30-March -2009 Updated: 16-November -2009

Expected Results: Mutant = 190 bp Heterozygote = 235 bp and 190 bp Wild type = 235 bp

Gel Image

Image Image file View Full Size

Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR8038 - TAA GCC TGC CCA GAA GAC TC - Wild type Reverse oIMR8545 - AAA GTC GCT CTG AGT TGT TAT - Common oIMR9493 - AAA TCA GAG AGA TCC TCA TAA AGG - Mutant Forward

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 oIMR8545 0.60 1.00 uM 0.60 20 uM oIMR8038 0.60 1.00 uM 0.60 20 uM oIMR9493 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 62 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 009043 129S-Gt(ROSA)26Sortm2Tyj/J 009044 B6.129S4-Gt(ROSA)26Sortm2Tyj/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.