Stock Number: 007708 Strain Name: B6.129-Gt(ROSA)26Sortm1(HD*103Q)Xwy/J

Allele: HD*103Q Protocol Name: HD*103Q Method: Standard PCR Version: 2.0 Created: 23-June -2009 Updated: 24-September-2009

Notes: FML Buffer: 500 mM KCl, 100 mM Tris HCl pH 8.3, 15 mM MgCl2, 0.01% Gelatin Size on ABI3730. Expected Results: Mutant = 359 bp

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 7708-65-L 6-FAM AAG GCC TTC GAG TCC CTC AAA A - - 7708-65-R - GAG GTT GAG GAA GTT GAG GAG GTG - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 8.11 - 8.11 10 X FML Buffer 1.31 1.09 X 1.31 2.5 mM dNTP 1.04 0.22 mM 1.04 6.6 uM 7708-65-L with 0.24 0.13 uM 0.24 6.6 uM 7708-65-R 0.24 0.13 uM 0.24 5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06 DNA (crude prep diluted 1:1 in water) 1.00 - 1.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 65 35 sec - 4 72 30 sec repeat steps 2-4 for 38 cycles 5 72 7 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 007708 B6.129-Gt(ROSA)26Sortm1(HD*103Q)Xwy/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.