Stock Number: 008600 Strain Name: B6.129P2-Gt(ROSA)26Sortm1(tTA)Roos/J

Allele: Gt(ROSA)26Sortm1(tTA)Roos Protocol Name: Gt(ROSA)26Sortm1(tTA)Roos Method: Standard PCR Version: 1.1 Created: 24-February -2009 Updated: 24-September-2009

Expected Results: Mutant = 224 bp Heterozygote = 297 bp and 224 bp Wild type = 297 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 oIMR9020 - AAG GGA GCT GCA GTG GAG TA - Common oIMR9021 - CCG AAA ATC TGT GGG AAG TC - Wild type Reverse oIMR9022 - TCA TCA AGG AAA CCC TGG AC - 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 oIMR9020 0.60 1.00 uM 0.60 20 uM oIMR9021 0.60 1.00 uM 0.60 20 uM oIMR9022 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 61 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 008600 B6.129P2-Gt(ROSA)26Sortm1(tTA)Roos/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.