Stock Number: 003275 Strain Name: STOCK Tg(tetL)1Bjd/J

Allele: Tg(tetL)1Bjd Protocol Name: Luciferase: Tg(tetNZL)2Bjd, Tg(tetL)1Bjd, Tg(Fos-luc)1Rnd Method: Standard PCR Version: 1.1 Created: 03-November -2000 Updated: 24-September-2009

Expected Results: TG = 465 bp internal control = ~200 bp

Gel Image

Image Image file View Full Size

Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0960 - TCC TCT GAC ACA TAA TTC GCC - Transgene oIMR0961 - GTT GTT CCA TTC CAT CAC GG - 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 3.16 - 3.16 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 oIMR0960 0.80 1.33 uM 0.80 20 uM oIMR0961 0.80 1.33 uM 0.80 20 oIMR8744 0.20 0.33 0.20 20 uM oIMR8745 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 0.06 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 60 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 003274 STOCK Tg(tetNZL)2Bjd/J 003275 STOCK Tg(tetL)1Bjd/J 003479 B6.C3-Tg(Fos-luc)1Rnd/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.