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.
Mut= 195 bp
Wt= 193 bp
Wt Sequence:
CTGGCTTCTGAGGACCGCCCTGGGCCTGGGAGAATCCCTTCCCCCTCTTCCCTCGTGATCTGCAACTCCAGtctttctagaaGATGGGCGGG
AGTCTTCTGGGCAGGCTTAAAGGCTAACCTGGTGTGTGGGCGTTGTCCTGCAGGGGAATTGAACAGGTGTAAAATTGGAGGGACAAGACTTCCCACAGATT
Mutant Sequence:
CTGGCTTCTGAGGACCGCCCTGGGCCTGGGAGAATCCCTTCCCCCTCTTCCCTCGTGATCTGCAACTCCAGTCTTTCTAGtgGATCCCCCg
ggcTGCAGATctgTAGGGCGCAGTAGTCCAGGGTTTCCTTGATGATGTCATACTTATCCTGTCCCTTTTTTTTCCACAGCTCGCGGTTGAGGACAAACTCTTCG
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
21306 | CTG GCT TCT GAG GAC CG | Common | A | Gt(ROSA)26Sor | ||
21307 | CGA AGA GTT TGT CCT CAA CCG | Mutant Reverse | A | SA site | ||
21308 | Fluorophore-1 | ACC CTG GAC TAC TGC GCC C | Quencher-1 | MUT Probe | SA site | |
21309 | AAT CTG TGG GAA GTC TTG TCC | Wild type Reverse | A | Gt(ROSA)26Sor | ||
21310 | Fluorophore-2 | TAA CCT GGT GTG TGG GCG TTG T | Quencher-2 | WT Probe | Gt(ROSA)26Sor |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
21306 | 0.40 uM |
21307 | 0.40 uM |
21309 | 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 |
---|---|
003310 | 129S-Gt(ROSA)26Sortm1Sor/J |
007844 | 129S4/SvJae-Gt(ROSA)26Sortm2(FLP*)Sor/J |
007689 | 129S4/SvJaeSor-Gt(ROSA)26Sortm4(attB/attP)Sor/J |
009575 | B6(129S4)-Et(cre/ERT2)119Rdav/J |
012688 | B6(129S4)-Et(cre/ERT2)13866Rdav/J |
009582 | B6(129S4)-Et(cre/ERT2)1645Rdav/J |
009585 | B6(129S4)-Et(cre/ERT2)2047Rdav/J |
009574 | B6(129S4)-Et(cre/ERT2)21Rdav/J |
010688 | B6(129S4)-Et(cre/ERT2)6691Rdav/J |
010689 | B6(129S4)-Et(cre/ERT2)6959Rdav/J |
010690 | B6(129S4)-Et(cre/ERT2)7089Rdav/J |
010691 | B6(129S4)-Et(cre/ERT2)7149Rdav/J |
010692 | B6(129S4)-Et(cre/ERT2)7381Rdav/J |
010693 | B6(129S4)-Et(cre/ERT2)8120Rdav/J |
010694 | B6(129S4)-Et(cre/ERT2)8131Rdav/J |
010695 | B6(129S4)-Et(cre/ERT2)9699Rdav/J |
012690 | B6(129S4)-Et(icre/ERT2)14208Rdav/J |
012687 | B6(129S4)-Tg(SYN1-icre/mRFP1)9934Rdav/J |
013730 | B6(Cg)-Calb2tm2.1(cre/ERT2)Zjh/J |
012704 | B6(Cg)-Crhtm1(cre)Zjh/J |
010705 | B6(Cg)-Dlx5tm1(cre/ERT2)Zjh/J |
010776 | B6(Cg)-Lhx6tm1(cre/ERT2)Zjh/J |
010777 | B6(Cg)-Pvalbtm1(cre/ERT2)Zjh/J |
007708 | B6.129-Gt(ROSA)26Sortm1(HD*103Q)Xwy/J |
008877 | B6.129P2(129S4)-Hprt1tm12(Ple177-EGFP/cre)Ems/Mmjax |
009114 | B6.129P2(129S4)-Hprt1tm14(Ple103-EGFP/cre)Ems/Mmjax |
008709 | B6.129P2(129S4)-Hprt1tm9(Ple178-EGFP/cre)Ems/Mmjax |
016933 | B6.129P2(Cg)-Cdh1tm1Cle/J |
011008 | B6.129P2(Cg)-Gt(ROSA)26Sortm1(tTA)Roos/J |
009115 | B6.129P2(Cg)-Hprt1tm15(Ple111-EGFP)Ems/Mmjax |
009348 | B6.129P2(Cg)-Hprt1tm17(Ple48-lacZ)Ems/Mmjax |
009118 | B6.129P2(Cg)-Hprt1tm18(Ple90-EGFP)Ems/Mmjax |
009596 | B6.129P2(Cg)-Hprt1tm33(Ple183-EGFP)Ems/Mmjax |
010709 | B6.129P2(Cg)-Hprt1tm44(Ple49-lacZ)Ems/Mmjax |
012580 | B6.129P2(Cg)-Hprt1tm59(Ple123-lacZ)Ems/Mmjax |
012581 | B6.129P2(Cg)-Hprt1tm62(Ple153-lacZ)Ems/Mmjax |
012583 | B6.129P2(Cg)-Hprt1tm68(Ple127-lacZ)Ems/Mmjax |
008707 | B6.129P2(Cg)-Hprt1tm7(Ple185-EGFP)Ems/Mmjax |
008708 | B6.129P2(Cg)-Hprt1tm8(Ple151-EGFP)Ems/Mmjax |
034676 | B6.129S(Cg)-Apoetm1(APOE*)Ljohn/J |
003474 | B6.129S4-Gt(ROSA)26Sortm1Sor/J |
012930 | B6.129S4-Gt(ROSA)26Sortm2(FLP*)Sor/J |
007743 | B6.129S4-Gt(ROSA)26Sortm3(phiC31*)Sor/J |
010671 | B6.129S4-Pkd1tm2Ggg/J |
009673 | B6.129S6(C)-Gt(ROSA)26Sortm3(HIF1A*)Kael/J |
032087 | B6.129S6(Cg)-Gt(ROSA)26Sortm1(DTA)Jpmb/J |
008466 | B6.129X1(Cg)-Shhtm6Amc/J |
021071 | B6.Cg-Gt(ROSA)26Sortm1(CAG-PA-GFP)Rmpl/J |
026293 | B6.Cg-Gt(ROSA)26Sortm1(LRRK2*R1441C)Djmo/J |
029784 | B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm1(tetO-U2AF1*S34F)Mjwa/J |
014588 | B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm6(tetO-MSI2)Jae/J |
017983 | B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm9(tetO-Dnmt3b_i1)Jae/J |
014602 | B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm1(tetO-mCherry)Eggn/J |
023749 | B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae Tg(tetO-Pou5f1,-Sox2,-Klf4,-Myc)1Srn/J |
006965 | B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae/J |
014536 | B6.Cg-Hprt1tm75(Ple143-lacZ)Ems/Mmjax |
016868 | B6.Cg-Ssttm1.2(tTA2)Hze/J |
021070 | B6.Cg-Tg(Thy1-PA-GFP)6Rmpl/J |
010527 | B6;129-Gt(ROSA)26Sortm1(DTA)Mrc/J |
016262 | B6;129-Gt(ROSA)26Sortm1(Foxo1/GFP)Jke/J |
017962 | B6;129-Gt(ROSA)26Sortm1(RAC1*)Jkis/J |
021025 | B6;129-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm1(tetO-cre)Haho/J |
006911 | B6;129-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm2(tetO-Pou5f1)Jae/J |
008883 | B6;129-Gt(ROSA)26Sortm1(SNCA*A53T)Djmo/TmdJ |
008889 | B6;129-Gt(ROSA)26Sortm2(SNCA*119)Djmo/TmdJ |
008886 | B6;129-Gt(ROSA)26Sortm3(SNCA*E46K)Djmo/TmdJ |
021039 | B6;129-Gt(ROSA)26Sortm5(CAG-Sun1/sfGFP)Nat/J |
023843 | B6;129-Gt(ROSA)26Sortm6(Fzd6)Nat/J |
026007 | B6;129-Gt(ROSA)26Sortm8(cre/Esr1*)Nat/J |
022516 | B6;129S-Gt(ROSA)26Sortm1(Cdkn1c)Jfpa/J |
012373 | B6;129S-Hoxb1tm1(cre)Og/J |
014592 | B6;129S4-Col1a1tm1(tetO-mCherry)Eggn/J |
011001 | B6;129S4-Col1a1tm1(tetO-Pou5f1,-Klf4,-Sox2,-Myc)Hoch/J |
012464 | B6;129S4-Foxd1tm2(GFP/cre/ERT2)Amc/J |
029415 | B6;129S4-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm1(tetO-cas9)Sho/J |
016836 | B6;129S4-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm7(tetO-HIST1H2BJ/GFP)Jae/J |
003309 | B6;129S4-Gt(ROSA)26Sortm1Sor/J |
009576 | B6;129S4-Et(cre/ERT2)278Rdav/J |
017738 | B6;SJL-Tg(Foxl1-cre)1Khk/J |
019101 | B6N.129S4(B6)-Gt(ROSA)26Sortm1Sor/CjDswJ |
019013 | B6N.129S6(Cg)-Gt(ROSA)26Sortm2(EGFP/cre)Alj/J |
008603 | C.129P2(B6)-Gt(ROSA)26Sortm1(tTA)Roos/J |
002955 | C.129S7-Gt(ROSA)26Sor/J |
007900 | C57BL/6-Gt(ROSA)26Sortm1(HBEGF)Awai/J |
016097 | C57BL/6-Tg(Car1-cre)5Flt/J |
008040 | CBy.B6-Gt(ROSA)26Sortm1(HBEGF)Awai/J |
026863 | D2.129S4(B6)-Gt(ROSA)26Sortm1Sor/SjJ |
011034 | FVB(Cg)-Tg(Ghrhr-cre)3242Lsk/J |
006954 | FVB-Tg(Ddx4-cre)1Dcas/J |
009427 | FVB.129S4(B6)-Gt(ROSA)26Sortm1Sor/J |
016977 | FVB.129S6-Gt(ROSA)26Sortm1(Pik3ca*H1047R)Egan/J |
016603 | NOD.B6-Gt(ROSA)26Sortm1(HBEGF)Awai/DvsJ |
010910 | STOCK Corttm1(cre)Zjh/J |
010702 | STOCK Gad2tm1(cre/ERT2)Zjh/J |
010802 | STOCK Gad2tm2(cre)Zjh/J |
006331 | STOCK Gt(ROSA)26Sortm1(DTA)Jpmb/J |
022793 | STOCK Gt(ROSA)26Sortm1(LRRK2*R1441C)Djmo/J |
008159 | STOCK Gt(ROSA)26Sortm1(Notch1)Dam/J |
034693 | STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Hprt1tm1(tetO-KMT2A/MLLT3)Sguo/J |
011004 | STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm3(tetO-Pou5f1,-Sox2,-Klf4,-Myc)Jae/J |
011011 | STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm4(tetO-Pou5f1,-Sox2,-Klf4,-Myc)Jae/J |
011013 | STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm5(tetO-Pou5f1,-Klf4,-Myc)Jae/J |
008600 | STOCK Gt(ROSA)26Sortm1(tTA)Roos/J |
025980 | STOCK Gt(ROSA)26Sortm1.1(birA)Mejr Ep300tm1.1Wtp/Mmjax |
025983 | STOCK Gt(ROSA)26Sortm1.1(birA)Mejr Mef2ctm1.1Wtp/Mmjax |
025978 | STOCK Gt(ROSA)26Sortm1.1(birA)Mejr Nkx2-5tm1.1Wtp/Mmjax |
025977 | STOCK Gt(ROSA)26Sortm1.1(birA)Mejr Zfpm2tm1.1Wtp /Mmjax |
018903 | STOCK Gt(ROSA)26Sortm2(EGFP/cre)Alj/J |
009674 | STOCK Gt(ROSA)26Sortm4(HIF2A*)Kael/J |
009349 | STOCK Hprt1tm31(Ple67-EGFP)Ems/Mmjax |
009594 | STOCK Hprt1tm32(Ple112-EGFP)Ems/Mmjax |
010707 | STOCK Hprt1tm37(lacZ)Ems/Mmjax |
012335 | STOCK Hprt1tm50(Ple55-lacZ)Ems/Mmjax |
012353 | STOCK Hprt1tm65(Ple53-lacZ)Ems/Mmjax |
012584 | STOCK Hprt1tm69(Ple134-lacZ)Ems/Mmjax |
016117 | STOCK Nogtm1.1Rmh/J |
006953 | STOCK Notch1tm3(cre)Rko/J |
028560 | STOCK Sncatm1.2Vlb Gt(ROSA)26Sortm1Sho/J |
014581 | STOCK Trpm8tm1Apat/J |
010908 | STOCK Viptm1(cre)Zjh/J |
010911 | STOCK Wt1tm1(EGFP/cre)Wtp/J |
012691 | STOCK Et(icre/ERT2)14374Rdav/J |
012692 | STOCK Et(icre/ERT2)14602Rdav/J |
012693 | STOCK Et(icre/ERT2)14624Rdav/J |
007684 | STOCK Tg(Atoh1-cre/Esr1*)14Fsh/J |
008852 | STOCK Tg(En2-cre)22Alj/J |
023965 | STOCK Tg(Krt17-EGFP)#Cou/J |
008847 | STOCK Tg(Msx2-cre/ERT)27Alj/J |
014158 | STOCK Tg(Pax4-cre)1Dam/J |
009606 | STOCK Tg(Six2-EGFP/cre)1Amc/J |
130 strains use this protocol |