Stock Number: 006235 Strain Name: B6.Cg-Tg(SFTPC-rtTA)5Jaw/J

Allele: Generic tTA Protocol Name: Tg(tTA) Method: Standard PCR Version: 2.1 Created: 28-March -2003 Updated: 13-October -2010

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene=450 bp Internal positive 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 oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control Forward oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control Reverse oIMR8746 - CGC TGT GGG GCA TTT TAC TTT AG - Transgene Forward oIMR8747 - CAT GTC CAG ATC GAA ATC GTC - Transgene Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.99 - 3.99 10 X AB PCR BufferII 0.96 0.80 X 0.96 25 mM MgCl2 1.20 2.50 mM 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 20 ul oIMR8744 0.30 0.50 ul 0.30 20 uM oIMR8745 0.30 0.50 uM 0.30 20 uM oIMR8746 0.30 0.50 uM 0.30 20 uM oIMR8747 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 3 min - 2 94 30 sec - 3 57 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 002618 B6.Cg-Tg(MMTVtTA)1Mam/J 002709 B6;C3-Tg(TettTALuc)1Dgs/J 003010 B6;CBA-Tg(Camk2a-tTA)1Mmay/J 003170 FVB.Cg-Tg(Myh6-tTA)6Smbf/J 003271 STOCK Tg(CMV-tTA)3Bjd/J 003272 STOCK Tg(Cebpb-tTA)5Bjd/J 003273 STOCK Tg(CMV-rtTA)4Bjd/J 003563 B6.Cg-Tg(Cebpb-tTA)5Bjd/J 003763 B6.Cg-Tg(Eno2tTA)5030Nes/J 003767 B6.Cg-Tg(Eno2tTA)5021Nes/J 004127 FVB-Tg(Nes-rtTA)306Rvs/J 004937 NOD.Cg-Tg(Ins2-tTA)1Doi/DoiJ 005493 STOCK Tg(Tek-rtTA,TRE-lacZ)1425Tpr/J 005625 FVB-Tg(Pcp2-tTA)3Horr/J 005734 NOD/Lt-Tg(Ins2-rtTA)1Ach/AchJ 005942 FVB/N-Tg(Pf4-tTA/VP16)42Kra/J 005964 B6.Cg-Tg(GFAP-tTA)110Pop/J 006209 FVB.Cg-Tg(Tal1-tTA)19Dgt/J 006235 B6.Cg-Tg(SFTPC-rtTA)5Jaw/J 006875 FVB/N-Tg(Tagln-rtTA)E1Jwst/J 006999 STOCK Dbttm1Geh Tg(Cebpb-tTA)5Bjd Tg(tetO-DBT)A1Geh/J 007004 B6.Cg-Tg(Camk2a-tTA)1Mmay/DboJ 007678 B6;SJL-Tg(KRT14-rtTA)208Jek/J 012433 B6;C3-Tg(ACTA1-rtTA,tetO-cre)102Monk/J 016146 STOCK Tg(SFTPC-rtTA)2Jaw/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.