Stock Number: 003380 Strain Name: B6.FVB-Tg(C3-1-TAg)cJeg/J

Allele: Tg(C3-1-TAg)cJeg Protocol Name: TAg, SV Method: Standard PCR Version: 2.1 Created: 15-July -2008 Updated: 24-September-2009

Expected Results: Internal Control = ~200 bp TG = ~500 bp

Gel Image

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Gel Information: Separate digested products on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0068 - CAG AGC AGA ATT GTG GAG TGG - Transgene oIMR0069 - GGA CAA ACC ACA ACT AGA ATG CAG TG - Transgene oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control Forward oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.65 - 3.65 10 X PCR Buffer II (ABI) 0.70 0.58 X 0.70 25 mM MgCl2 1.20 2.50 mM 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR0068 0.60 1.00 uM 0.60 20 uM oIMR0069 0.60 1.00 uM 0.60 20 oIMR8744 0.30 0.50 0.30 20 uM oIMR8745 0.30 0.50 uM 0.30 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 1.5 min - 2 94 20 sec - 3 64 30 sec* *-0.5 C per cycle 4 72 35 sec repeat steps 2-4 for 12 cycles 5 94 20 sec - 6 58 30 sec - 7 72 35 sec repeat steps 5-7 for 25 cycles 8 72 2 min - 9 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 002033 NOD/ShiLt-Tg(RipTAg)1Lt/J 002233 C57BL/6J-Tg(SV)7Bri/J 002380 NOD.Cg-Tg(Ins2-TAg)1Lt Prkdcscid/DvsJ 003188 C57BL/6J-Tg(WapTAg)1Knw 003189 C57BL/6J-Tg(WAPTAg)3Knw/J 003380 B6.FVB-Tg(C3-1-TAg)cJeg/J 003381 FVB-Tg(C3-1-TAg)cJeg/J 003382 B10.D2-Tg(C3-1-TAg)cJeg/J 003445 C57BL/6J-Tg(Amy1TAg)354Knw/J 003446 C57BL/6J-Tg(Amy1TAg)501Knw/J 003476 C57BL/6J-Tg(SV)427Bri/J 003477 C57BL/6J-Tg(SV)419Bri/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.