Stock Number: 005769 Strain Name: B6.129P2-Htr7tm1Dgen/J

Allele: Htr7tm1Dgen Protocol Name: Htr7tm1Dgen Method: Standard PCR Version: 1.1 Created: 05-September-2008 Updated: 11-January -2010

Expected Results: Mutant = 494 bp Heterozygote = 307 bp and 494 bp Wild type = 307 bp

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

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type moIMR0012 - GGG TGG GAT TAG ATA AAT GCC TGC TCT - - oIMR4384 - CAT GTC GGT CAT GCT GTT CAT GTA C - - oIMR4385 - TAA AGG GCC TTG CTG TGG ACA AGA G - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.94 - 4.94 10 X AB PCR BufferII 1.00 1.00 X 1.00 50 mM MgCl2 0.30 1.50 mM 0.30 2.5 mM dNTP 0.96 0.24 mM 0.96 20 uM oIMR4384 0.25 0.50 uM 0.25 20 uM oIMR4385 0.25 0.50 uM 0.25 20 uM moIMR0012 0.25 0.50 uM 0.25 5 U/ul TEMPase II Taq Pol. 0.05 0.03 U/ul 0.05 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 95 7 min - 2 94 30 sec - 3 60 30 sec - 4 68 1.5 min repeat steps 2-4 for 35 cycles 5 68 7 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 005769 B6.129P2-Htr7tm1Dgen/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.