Stock Number: 008232 Strain Name: FVB/N-Tg(Ins2-IAPP)RHFSoel/J

Allele: Tg(Ins2-IAPP)RHFSoel Protocol Name: Tg(Ins2-IAPP)RHFSoel Method: Standard PCR Version: 2.1 Created: 15-September-2008 Updated: 24-September-2009

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = ~954bp Internal positive control = 324 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 oIMR7338 - CTA GGC CAC AGA ATT GAA AGA TCT - Internal Positive Control Forward oIMR7339 - GTA GGT GGA AAT TCT AGC ATC ATC C - Internal Positive Control Reverse oIMR8417 - GTC ATG TGC ACC TAA AGG GGC AAG TAA TTC A - Transgene Forward oIMR8418 - CGA GTG GGC TAT GGG TTT GT - Transgene Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.46 - 3.46 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 oIMR8417 0.60 1.00 uM 0.60 20 uM oIMR8418 0.60 1.00 uM 0.60 20 uM oIMR7338 0.25 0.42 uM 0.25 20 uM oIMR7339 0.25 0.42 uM 0.25 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 63 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 008232 FVB/N-Tg(Ins2-IAPP)RHFSoel/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.