Stock Number: 003393 Strain Name: B6;SJL-Tg(aP2-SREBF1c)9884Reh/J

Allele: Tg(aP2-SREBF1c)9884Reh Protocol Name: Tg(aP2-SREBF1c) Method: Standard PCR Version: 1.0 Created: 17-July -2009 Updated: 24-September-2009

Expected Results: Transgene = 151 bp 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 oIMR0135 - CAT CCC TGT GAC CCC TCC - Transgene Forward oIMR0136 - CTC CAA ACC ACC CCC CTC - Transgene Reverse 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

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.83 - 3.83 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 oIMR0135 0.48 0.80 uM 0.48 20 uM oIMR0136 0.48 0.80 uM 0.48 20 uM oIMR7338 0.20 0.33 uM 0.20 20 uM oIMR7339 0.20 0.33 uM 0.20 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul Taq DNA Polymerase 0.03 0.01 U/ul 0.03 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 20 sec - 3 57 30 sec - 4 72 20 sec 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 003393 B6;SJL-Tg(aP2-SREBF1c)9884Reh/J 613393 B6;SJL-Tg(aP2-SREBF1c)9884Reh/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.