Stock Number: 007042 Strain Name: D2.129P2(B6)-Nr5a1tm2Klp/EiJ

Allele: Nr5a1tm2Klp Protocol Name: Nr5a1tm2Klp Method: Standard PCR Version: 1.1 Created: 03-July -2008 Updated: 01-April -2010

Expected Results: Mutant = ~210 bp Heterozygote = 162 bp and ~210 bp Wild type = 162 bp

Gel Image

Image Image file View Full Size

Gel Information: Separated by gel electrophoresis on a 3.0% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR7290 - AGA CAA GTG CAC CCC ATT TC - Forward oIMR7291 - ACC ATC ACC AAC CGC TAA AC - Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.96 - 3.96 10 X AB PCR BufferII 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 oIMR7290 0.60 1.00 uM 0.60 20 uM oIMR7291 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 60 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 007041 B6Ei.129P2-Nr5a1tm2Klp/EiJ 007042 D2.129P2(B6)-Nr5a1tm2Klp/EiJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.