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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/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 | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
17817 | ACA ACT ACC TGT TCT GCC G | Forward | A | |||
17818 | GCC TCA AAG ATC CCT TCC AG | Reverse | A | |||
17819 | Fluorophore-1 | TTC TGC CAC CTC CCA ACT GTC C | Quencher-1 | Tg Probe | ||
oIMR1544 | CAC GTG GGC TCC AGC ATT | Internal Positive Control Forward | A | |||
oIMR3580 | TCA CCA GTC ATT TCT GCC TTT G | Internal Positive Control Reverse | A | |||
TmoIMR0105 | Fluorophore-2 | CCA ATG GTC GGG CAC TGC TCA A | Quencher-2 | IC Probe |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa Probe Fast QPCR | 1.00 X |
17817 | 0.40 uM |
17818 | 0.40 uM |
oIMR1544 | 0.40 uM |
oIMR3580 | 0.40 uM |
Tg Probe | 0.15 uM |
IC Probe | 0.15 uM |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 95.0 | -- | |
2 | 95.0 | -- | |
3 | 60.0 | -- | repeat steps 2-3 for 40 cycles |
Stock Number | Strain Name |
---|---|
035764 | B6(129)-Jchainem1(icre/ERT2)Deep/J |
035435 | B6(129S4)-Xcr1tm1.1(cre)Kmm/J |
010696 | B6(129S4)-Et(icre/ERT2)10596Rdav/J |
010697 | B6(129S4)-Et(icre/ERT2)10727Rdav/J |
037836 | B6(FVB)-Il1bem1Mora/J |
038129 | B6.Cg-Ascl3tm2.1(EGFP/icre/ERT2)Ovi/J |
038342 | B6.Cg-Gas1em1(icre/ERT2)Nono/J |
023691 | B6.Cg-Hprt1tm346a(Ple264-icre/ERT2)Ems/Mmjax |
026516 | B6.Cg-Tg(Avil-icre/ERT2)AJwo/J |
008520 | B6.Cg-Tg(CD2-icre)4Kio/J |
036935 | B6.FVB(Cg)-Tp(X)1SoffTg(Myh11-icre/ERT2)1Soff/ZjngJ |
019079 | B6.FVB-Tg(Myh11-icre/ERT2)1Soff/J |
017490 | B6.FVB-Tg(Stra8-icre)1Reb/LguJ |
030663 | B6;129S-Gcgtm1.1(icre)Gkg/J |
025809 | B6;CBA-Tg(Fgfr3-icre/ERT2)4-2Wdr/J |
024768 | B6;CBA-Tg(Upk2-icre/ERT2)1Ccc/J |
031645 | B6;D2-Tn(pb-CAG-cas9*,-EGFP)1Yangh/J |
028547 | B6;SJL-Tg(Cyp17a1-icre)AJako/J |
036235 | B6N.Cg-Tg(Cxcl12-icre/ERT2)L289Nono/J |
035229 | C57BL/6-Ccr2em1(icre/ERT2)Peng/J |
025124 | C57BL/6-Tg(Adipoq-icre/ERT2)1Soff/J |
038197 | C57BL/6-Tg(Nmur1-iCre,-eGFP)1Dart/J |
016583 | C57BL/6-Tg(Slc6a3-icre/ERT2)2Gloss/J |
030676 | C57BL/6N-Tg(Tg-cre/ERT2)1Kero/J |
019098 | FVB-Tg(Csf1r-cre/Esr1*)1Jwp/J |
021024 | FVB-Tg(Csf1r-icre)1Jwp/J |
037019 | STOCK Foxp3tm4(icre)Tch/Mmjax |
034261 | STOCK Ighg3tm1.1(icre)Gbrt/J |
037139 | STOCK Myl1tm1(cre)Sjb Atg7tm1Tchi Gaatm1Rabn/PuroMmjax |
008208 | STOCK Tg(Stra8-icre)1Reb/J |
030061 | STOCK Tg(Tbx18-icre)3Fech/J |
016584 | STOCK Tg(Tph2-icre/ERT2)6Gloss/J |
32 strains use this protocol |