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This assay will NOT distinguish hemizygous from homozygous
Transgene = 89 bp
Internal positive control = 74 bp
Tg Sequence:
ATGCTCCTGTCTGTGTGCAGATCCTGGGCTGCCTGGTGCAAGCTGAACAACAGGAAATGGTTCCCTGCTGAACCTGAGGATGTGAGGGA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
22926 | TCC CTC ACA TCC TCA GGT TC | Transgene Reverse | A | |||
41665 | ATG CTC CTG TCT GTG TGC AG | Transgene Forward | A | |||
41666 | Fluorophore-1 | TGC CTG GTG CAA GCT GA | 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 |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
22926 | 0.40 uM |
41665 | 0.40 uM |
oIMR1544 | 0.40 uM |
oIMR3580 | 0.40 uM |
Wt Probe | 0.15 uM |
Mutant Probe | 0.15 uM |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 95.0 | -- | |
2 | 95.0 | -- | |
3 | 60.0 | -- | |
4 | -- | repeat steps 2-3 for 40 cycles | |
5 | 40.0 | -- | Forever |
Stock Number | Strain Name |
---|---|
035764 | B6(129)-Jchainem1(icre/ERT2)Deep/J |
035435 | B6(129S4)-Xcr1tm1.1(cre)Kmm/J |
009587 | B6(129S4)-Et(icre)1402Rdav/J |
009588 | B6(129S4)-Et(icre)1470Rdav/J |
009589 | B6(129S4)-Et(icre)1555Rdav/J |
009586 | B6(129S4)-Et(icre)754Rdav/J |
010696 | B6(129S4)-Et(icre/ERT2)10596Rdav/J |
010697 | B6(129S4)-Et(icre/ERT2)10727Rdav/J |
012689 | B6(129S4)-Et(icre/ERT2)14163Rdav/J |
012690 | B6(129S4)-Et(icre/ERT2)14208Rdav/J |
012694 | B6(129S4)-Et(icre/ERT2)14915Rdav/J |
012687 | B6(129S4)-Tg(SYN1-icre/mRFP1)9934Rdav/J |
037836 | B6(FVB)-Il1bem1Mora/J |
038129 | B6.Cg-Ascl3tm2.1(icre/ESR1*)Ovi/J |
008610 | B6.Cg-Commd10Tg(Vav1-icre)A2Kio/J |
038342 | B6.Cg-Gas1em1(icre/ERT2)Nono/J |
023677 | B6.Cg-Hprt1tm332(Ple272-icre/ERT2)Ems/Mmjax |
023690 | B6.Cg-Hprt1tm345a(icre/ERT2)Ems/Mmjax |
023691 | B6.Cg-Hprt1tm346a(Ple264-icre/ERT2)Ems/Mmjax |
023692 | B6.Cg-Hprt1tm347a(Ple155-icre/ERT2)Ems/Mmjax |
023693 | B6.Cg-Hprt1tm348a(Ple265-icre/ERT2)Ems/Mmjax |
023694 | B6.Cg-Hprt1tm349a(hs671-icre/ERT2)Ems/Mmjax |
023695 | B6.Cg-Hprt1tm350a(Ple232-icre/ERT2)Ems/Mmjax |
023696 | B6.Cg-Hprt1tm351a(Ple198-icre/ERT2)Ems/Mmjax |
023697 | B6.Cg-Hprt1tm352a(Hspa1a-icre/ERT2)Ems/Mmjax |
023698 | B6.Cg-Hprt1tm353(Ple274-icre/ERT2)Ems/Mmjax |
023699 | B6.Cg-Hprt1tm354(Ple283-icre/ERT2)Ems/Mmjax |
023705 | B6.Cg-Hprt1tm360a(Ple253-icre/ERT2)Ems/Mmjax |
023706 | B6.Cg-Hprt1tm361a(Ple267-icre/ERT2)Ems/Mmjax |
023708 | B6.Cg-Hprt1tm363a(hs1218-icre/ERT2)Ems/Mmjax |
023709 | B6.Cg-Hprt1tm364a(CAG-icre/ERT2)Ems/Mmjax |
023710 | B6.Cg-Hprt1tm365a(Ple266-icre/ERT2)Ems/Mmjax |
023711 | B6.Cg-Hprt1tm366a(Ple67-icre/ERT2)Ems/Mmjax |
023712 | B6.Cg-Hprt1tm367a(Ple251-icre/ERT2)Ems/Mmjax |
038823 | B6.Cg-Myh11em1(icre)Jifz/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 |
029231 | B6;129-Bhlha15tm4Skz/J |
030663 | B6;129S-Gcgtm1.1(icre)Gkg/J |
038989 | B6;C3-Oxtrem4(icre/ERT2)Yinn/2J |
025809 | B6;CBA-Tg(Fgfr3-icre/ERT2)4-2Wdr/J |
026555 | B6;CBA-Tg(Lhx6-icre)1Kess/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 |
018968 | B6N.Cg-Commd10Tg(Vav1-icre)A2Kio/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 |
038650 | C57BL/6-Tg(Crhr1-icre,-tdTomato)#Njus/J |
038197 | C57BL/6-Tg(Nmur1-iCre,-eGFP)1Dart/J |
016582 | C57BL/6N-Tg(Slc32a1-icre/ERT2)3Gloss/J |
030676 | C57BL/6N-Tg(Tg-cre/ERT2)1Kero/J |
021024 | FVB-Tg(Csf1r-icre)1Jwp/J |
039016 | NOD.Cg-H2b Commd10Tg(Vav1-icre)A2Kio/JkraJ |
008694 | NOD/ShiLt-Tg(Foxp3-EGFP/cre)1cJbs/J |
037019 | STOCK Foxp3tm4(icre)Tch/Mmjax |
034261 | STOCK Ighg3tm1.1(icre)Gbrt/J |
037139 | STOCK Myl1tm1(cre)Sjb Atg7tm1Tchi Gaatm1Rabn/PuroMmjax |
012691 | STOCK Et(icre/ERT2)14374Rdav/J |
012692 | STOCK Et(icre/ERT2)14602Rdav/J |
012693 | STOCK Et(icre/ERT2)14624Rdav/J |
027798 | STOCK Tg(Nkx6-2-icre)1Kess/SshiJ |
018147 | STOCK Tg(Slc17a8-icre)1Edw/SealJ |
008208 | STOCK Tg(Stra8-icre)1Reb/J |
016584 | STOCK Tg(Tph2-icre/ERT2)6Gloss/J |
68 strains use this protocol |