Stock Number: 006336 Strain Name: B6.129-Selplgtm1Rpmc/J

Allele: Selplgtm1Rpmc Protocol Name: Selplgtm1Rpmc Method: Standard PCR Version: 1.1 Created: 25-June -2008 Updated: 24-September-2009

Expected Results: Mutant = 365 bp Heterozygote = 202 bp and 365 bp Wild type = 202 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 oIMR7915 - CGG TTG GCA TTA TTT GTG TG - Common oIMR7916 - CCA AGA TGG TCA GCA GCA C - Wild type Reverse oIMR8162 - TGG ATG TGG AAT GTG TGC GAG - Mutant Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.12 - 3.12 10 X PCR Buffer II (ABI) 1.20 1.00 X 1.20 25 mM MgCl2 1.20 2.50 mM 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR7915 0.60 1.00 uM 0.60 20 uM oIMR7916 0.60 1.00 uM 0.60 20 uM oIMR8162 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 61 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 006336 B6.129-Selplgtm1Rpmc/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.