Stock Number: 007177 Strain Name: B6.129P2-Mecp2tm1Bird/J

Allele: Mecp2tm1Bird Protocol Name: Mecp2tm1Bird 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 = 480 bp Heterozygote = 480 bp and 416 bp Wild type = 416 bp

Gel Image

Image Image file View Full Size

Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6833 - TGG TAA AGA CCC ATG TGA CCC AAG - Common oIMR6834 - GGC TTG CCA CAT GAC AAG AC - - oIMR6835 - TCC ACC TAG CCT GCC TGT ACT TTG - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.96 - 3.96 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 oIMR6833 0.60 1.00 uM 0.60 20 uM oIMR6834 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 69 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 006847 B6;129P2-Mecp2tm1Bird/J 007177 B6.129P2-Mecp2tm1Bird/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.