Stock Number: 012361 Strain Name: C57BL/6-Gt(ROSA)26Sortm9(Rac1*,EGFP)Rsky/J

Allele: Gt(ROSA)26Sortm9(Rac1*,EGFP)Rsky Protocol Name: Gt(ROSA)26Sortm9(Rac1*,EGFP)Rsky STD PCR Method: Standard PCR Version: 1.0 Created: 06-May -2010 Updated: 15-July -2010

Expected Results: Mutant = 421 bp Heterozygote = 421 bp and 297 bp Wild type = 297 bp

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR8436 - TCG CCT TCT TGA CGA GTT CT - Mutant oIMR9020 - AAG GGA GCT GCA GTG GAG TA - Common oIMR9021 - CCG AAA ATC TGT GGG AAG TC - Wild type Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.32 - 4.32 2.5 mM dNTP 0.96 0.20 mM 0.96 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 25 mM MgCl2 0.00 0.00 mM 0.00 DNA 2.00 - 2.00 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 20 uM oIMR9020 0.60 1.00 uM 0.60 20 uM oIMR9021 0.60 1.00 uM 0.60 20 uM oIMR8436 0.60 1.00 uM 0.60 5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 63 30 sec - 4 72 30 sec - 5 - - Go to step 2-4, 35 times 6 72 2 min - 7 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 012361 C57BL/6-Gt(ROSA)26Sortm9(Rac1*,EGFP)Rsky/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.