Stock Number: 012893 Strain Name: B6.Cg-Tg(S100a4-EGFP)M1Egn/YunkJ

Allele: Tg(S100a4-EGFP)M1Egn Protocol Name: Tg(S100a4-EGFP) Method: Standard PCR Version: 1.0 Created: 16-August -2010 Updated: 10-February -2011

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = ~180 bp Internal Positive Control = 324 bp

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 10889 - GTG ATT TGG GTC ATG CTC AG - Transgene Forward 10890 - GAA CAG CTC CTC GCC CTT GC - 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) 2.5 mM dNTP 0.96 0.20 mM 0.96 DNA 2.00 - 2.00 ddH2O 3.72 - 3.72 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 20 uM 10889 0.60 1.00 uM 0.60 20 uM 10890 0.60 1.00 uM 0.60 5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06 20 uM oIMR7338 0.60 1.00 uM 0.60 20 uM oIMR7339 0.60 1.00 uM 0.60

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30sec - 3 68 30sec - 4 72 30sec - 5 - - Go to step 2-4, 35 times 6 72 2 min - 7 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 012893 B6.Cg-Tg(S100a4-EGFP)M1Egn/YunkJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.