Stock Number: 002328 Strain Name: C.129S2-Plautm1Mlg/J

Allele: Plautm1Mlg Protocol Name: Plautm1 Method: Standard PCR Version: 3.0 Created: 28-August -2009 Updated: 24-September-2009

Expected Results: Mutant = 337 bp Heterozygote = 153 bp and 337 bp Wild type = 153 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 oIMR0432 - TCT GGA GGA CCG CTT ATC TG - Wild type Forward oIMR0433 - CTC TTC TCC AAT GTG GGA TTG - Common oIMR2060 - CAC GAG ACT AGT GAG ACG TG - Mutant Forward

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 oIMR2060 0.60 1.00 uM 0.60 20 uM oIMR0432 0.60 1.00 uM 0.60 20 uM oIMR0433 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 52 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 002328 C.129S2-Plautm1Mlg/J 002329 FVB.129S2-Plautm1Mlg/J 002509 B6.129S2-Plautm1Mlg/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.