Stock Number: 002033 Strain Name: NOD/ShiLt-Tg(RipTAg)1Lt/J

Allele: Tg(RipTAg)1Lt Protocol Name: TAg Method: QPCR Version: 1.0 Created: 27-August -2009 Updated: 24-September-2009

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1544 - CAC GTG GGC TCC AGC ATT - Internal Positive Control Forward oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - Internal Positive Control Reverse oIMR9161 - CAT GAA CAG ACT GTG AGG ACT GAG - - oIMR9162 - GGA AGA CTC AGG GCA TGA AAC - - TmoIMR0107 JOE NHS Ester CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 1 Probe TmoIMR0133 6-FAM CCT GAA ATG AGC CTT GGG ACT GTG A Black Hole Quencher 1 -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 2 X Universal Master Mix 12.50 1.00 X 12.50 40 uM oIMR9161 0.25 0.40 uM 0.25 40 uM oIMR9162 0.25 0.40 uM 0.25 40 uM oIMR1544 0.25 0.40 uM 0.25 40 uM oIMR3580 0.25 0.40 uM 0.25 10 uM TmoIMR0133 0.75 0.30 uM 0.75 10 uM TmoIMR0107 0.75 0.30 uM 0.75 DNA 10.00 - 10.00

Cycling  Step # Temp °C Time Note 1 50 0 - 2 95 2 min - 3 95 15 sec - 4 60 1 min repeat steps 3-4 for 40 cycles

This genotyping protocol is used for the following strains:   Stock Number Strain Name 002033 NOD/ShiLt-Tg(RipTAg)1Lt/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.