Stock Number: 006940 Strain Name: C57BL/6-Cd22tm1Lam /J

Allele: Cd22tm1Lam Protocol Name: Cd22tm1Lam Method: Standard PCR Version: 1.1 Created: 09-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 620 bp Heterozygote = 286 bp and 620 bp Wild type = 286 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 oIMR8030 - CTC GTC TTA CCT TTT TGT TCC TCA - Wild type Forward oIMR8031 - AGT CCA GAG ACC ATC GGC AAG - Wild type Reverse oIMR8032 - GCG GGA AGG GAC TGG CTG CTA T - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.60 - 3.60 10 X PCR Buffer II (ABI) 0.96 0.80 X 0.96 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR8030 0.60 1.00 uM 0.60 20 uM oIMR8031 0.60 1.00 uM 0.60 20 uM oIMR8032 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 68 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 006940 C57BL/6-Cd22tm1Lam /J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.