Stock Number: 008532 Strain Name: B6;129-Thtm1(cre/Esr1)Nat/J

Allele: Thtm1(cre/ERT)Nat Protocol Name: Thtm1(cre/Esr1)Nat Method: Standard PCR Version: 1.1 Created: 18-September-2008 Updated: 13-June -2012

Expected Results: HET = 377 bp and 570 bp MUT = 570 bp WT = 377 bp

Gel Image

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

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR8703 - CGC ATA GAA ATT GCA TCA ACG CAT - Mutant Forward oIMR8704 - AGT TTG ACC CGT ACA CCC TGG CCA T - Wild type Forward oIMR8705 - GGA GAC CTT TCC TTC CTT TAT TGA GA - Common

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 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 oIMR8703 0.60 1.00 uM 0.60 20 uM oIMR8704 0.60 1.00 uM 0.60 20 uM oIMR8705 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 008532 B6;129-Thtm1(cre/Esr1)Nat/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.