Stock Number:004127
Strain Name:FVB-Tg(Nes-rtTA)306Rvs/J
Generic tTA
Tg(tTA)
QPCR
7.0
16-December -2011 29-January -2013
Notes:

Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler 480. Use an appropriate instrument-specific Fuorophore/Quencher combination.  The transgene genotype is determined by comparing ΔCt values of each unknown sample against known homozygous and hemizygous controls, using appropriate endogenous references.

Primer 9344 replaced with 13639, 3' Iowa Black with Zen
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
13734 6-FAM CGC ATG CCG ACG CGC TAG

Zen probe with Iowa Black

 -
13745 6-FAM CCA ATG GTC GGG CAC TGC TCA A

Zen probe with Iowa Black

 -
9342 - GGA CGA GCT CCA CTT AGA CG - -
9343 - CAA CAT GTC CAG ATC GAA ATC - -
oIMR1544 - CAC GTG GGC TCC AGC ATT - -
oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - -
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
40 uM 9343 0.26 0.83 uM 0.26
40 uM oIMR1544 0.13 0.42 uM 0.13
40 uM oIMR3580 0.13 0.42 uM 0.13
DNA 5.00 - 5.00
40 uM 9342 0.26 0.83 uM 0.26
2 X PerfeCTa qPCR SuperMix 6.25 1.00 X 6.25
ddH2O -0.26 - -0.26
5 uM 13734 0.38 0.15 uM 0.38
5 uM 13745 0.38 0.15 uM 0.38
Cycling 
Step # Temp °C Time Note
1 50 2 min -
2 95 10 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
002618 B6.Cg-Tg(MMTVtTA)1Mam/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
004127 FVB-Tg(Nes-rtTA)306Rvs/J
004937 NOD.Cg-Tg(Ins2-tTA)1Doi/DoiJ
005734 NOD/Lt-Tg(Ins2-rtTA)1Ach/AchJ
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
008250 STOCK Tg(Ins2-rtTA)2Efr/J
008755 STOCK Tg(Ins2-rtTA)2Efr Tg(teto-DTA)1Gfi/J
016145 STOCK Tg(Scgb1a1-rtTA)2Jaw/J
016146 STOCK Tg(SFTPC-rtTA)2Jaw/J
017519 STOCK Tg(KRT5-rtTA)T2D6Sgkd/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.