Stock Number: 007853 Strain Name: 129S/SvEv-Tg(Alb1-Ren)1Unc/CofJ

Allele: Tg(Alb1-Ren)1Unc Protocol Name: Tg(Alb1-Ren)1Unc Method: Standard PCR Version: 1.1 Created: 25-August -2008 Updated: 24-September-2009

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Mutant = 330 bp Heterozygote = 330 bp and 425 bp Wild type = 425 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 oIMR7647 - CAC AGA TTG TAA CTG CAA ATC TGT CG - Wild type Forward oIMR7648 - TGG GAT TCT AAC CCT GAG GAC C - Wild type Reverse oIMR7649 - GTT CTT CTG AGG GGA TCG GC - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 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 oIMR7647 0.60 1.00 uM 0.60 20 uM oIMR7648 0.60 1.00 uM 0.60 20 uM oIMR7649 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 64 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 007853 129S/SvEv-Tg(Alb1-Ren)1Unc/CofJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.