Stock Number: 006849 Strain Name: B6.129P2-Mecp2tm2Bird/J

Allele: Mecp2tm2Bird Protocol Name: Mecp2tm2Bird Method: Standard PCR Version: 1.1 Created: 02-July -2008 Updated: 24-September-2009

Notes: The Mecp2 gene is X-LINKED. Males with the targeted allele are hemizygous mutants Expected Results: Mutant = 222 bp Heterozygote = 222 bp and 379 bp Wild type = 379 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 oIMR7171 - AAC AGT GCC AGC TGC TCT TC - Common oIMR7172 - CTG TAT CCT TGG GTC AAG CTG - Wild type Reverse oIMR7415 - GCC AGA GGC CAC TTG TGT AG - Mutant

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 oIMR7171 0.60 1.00 uM 0.60 20 uM oIMR7172 0.60 1.00 uM 0.60 20 uM oIMR7415 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 67 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 006849 B6.129P2-Mecp2tm2Bird/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.