Stock Number: 007906 Strain Name: B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze/J

Allele: Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze Protocol Name: Gt(ROSA)26Sortm6(CAG-ZsGreen1) Method: Standard PCR Version: 3.0 Created: 21-May -2010 Updated: 21-May -2010

Expected Results: Mutant = 199 bp Heterozygote = 297 bp and 199 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 - Wild type Forward oIMR9021 - CCG AAA ATC TGT GGG AAG TC - Wild type Reverse oIMR9103 - GGC ATT AAA GCA GCG TAT CC - Mutant Reverse oIMR9104 - AAC CAG AAG TGG CAC CTG AC - Mutant Forward

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06 ddH2O 4.32 - 4.32 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR9020 0.45 0.75 uM 0.45 20 uM oIMR9021 0.45 0.75 uM 0.45 20 uM oIMR9104 0.45 0.75 uM 0.45 20 uM oIMR9103 0.45 0.75 uM 0.45 DNA 2.00 - 2.00 5 M Betaine 1.66 0.69 M 1.66

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 20 sec - 3 62 30 sec - 4 72 30 sec 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 007906 B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.