Stock Number: 008309 Strain Name: C57BL/6-Rag2tm1Cgn/J

Allele: Rag2tm1Cgn Protocol Name: Rag2tm1Cgn STD Method: Standard PCR Version: 2.0 Created: 12-July -2010 Updated: 12-July -2010

Expected Results: Mutant = 305 bp Heterozygote = 265 bp and 305 bp Wild type = 265 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 3.0% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR8330 - CAG CGC TCC TCC TGA TAC TC - Forward oIMR8331 - TGC ATT CCT AGA GCG TCC TT - Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.92 - 4.92 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR8330 0.60 1.00 uM 0.60 20 uM oIMR8331 0.60 1.00 uM 0.60 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06 DNA 2.00 0.00 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 58 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 008309 C57BL/6-Rag2tm1Cgn/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.