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The size of the mutant band can vary between different Gt(Rosa)26sor(CAG) strains.
>chr6:113025964-113026059 96bp CTTCCCTCGTGATCTGCAAC CAGGACAACGCCCACACA
Mutant= 140 bp
Wild Type = 96 bp
Fam=Mut
Hex=Wt
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
17062 | GTT ATG TAA CGC GGA ACT CCA | Mutant Reverse | A | |||
24500 | CAG GAC AAC GCC CAC ACA | Wild type Reverse | A | |||
36158 | CTT CCC TCG TGA TCT GCA AC | Common | A | |||
39816 | Fluorophore-1 | ACG GGG TCA TTA GTT CAT AGC CCA | Quencher-1 | MUT Probe | ||
40263 | Fluorophore-2 | CCA GTC TTT CTA GAA GAT GGG CGG | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
17062 | 0.40 uM |
24500 | 0.40 uM |
36158 | 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 |
---|---|
006053 | 129-Gt(ROSA)26Sortm1(CAG-EGFP)Luo/J |
006067 | 129-Gt(ROSA)26Sortm2(CAG-Dsred2/EGFP)Luo/J |
006041 | 129-Gt(ROSA)26Sortm3(CAG-EGFP/Dsred2)Luo/J |
007676 | B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J |
006071 | B6.129-Gt(ROSA)26Sortm1(CAG-EGFP)Luo/J |
006080 | B6.129-Gt(ROSA)26Sortm2(CAG-Dsred2/EGFP)Luo/J |
006075 | B6.129-Gt(ROSA)26Sortm3(CAG-EGFP/Dsred2)Luo/J |
022367 | B6.129S4-Gt(ROSA)26Sortm1(CAG-EGFP/Rpl10a,-birA)Wtp/J |
017626 | B6.Cg-Gt(ROSA)26Sortm1(CAG-GFP/Eif2c2)Zjh/J |
026294 | B6.Cg-Gt(ROSA)26Sortm2.2Ksvo/J |
017455 | B6;129-Gt(ROSA)26Sortm1(CAG-COP4*E123T*H134R,-tdTomato)Gfng/J |
021429 | B6;129-Gt(ROSA)26Sortm4(CAG-EGFP*)Nat/J |
021039 | B6;129-Gt(ROSA)26Sortm5(CAG-Sun1/sfGFP)Nat/J |
026006 | B6;129-Gt(ROSA)26Sortm7(CAG-tdTomato*)Nat/J |
026005 | B6;129-Gt(ROSA)26Sortm9(CAG-GFP*)Nat/J |
010523 | B6;129P2-Gt(ROSA)26Sortm1(CAG-ALPP)Fawa/J |
018385 | B6;129S-Gt(ROSA)26Sortm1(CAG-COX8A/Dendra2)Dcc/J |
024846 | B6;129S-Gt(ROSA)26Sortm2.1Ksvo/J |
023035 | B6;129S6-Gt(ROSA)26Sortm1(CAG-tdTomato*,-EGFP*)Ees/J |
036272 | B6;CBA-Gt(ROSA)26Sorem1(CAG-Ikzf2,-tdTomato)Zyliu/J |
026556 | B6N.129(B6J)-Gt(ROSA)26Sortm1(CAG-cas9*,-EGFP)Fezh/J |
026430 | B6N.129(Cg)-Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J |
023537 | B6N.129S6-Gt(ROSA)26Sortm1(CAG-tdTomato*,-EGFP*)Ees/J |
031928 | C57BL/6N-Gt(ROSA)26Sortm1(CAG-Il11)Cook/J |
026862 | D2.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/SjJ |
026481 | FVB.129(B6)-Gt(ROSA)26Sortm1(CAG-cas9*,-EGFP)Fezh/J |
026558 | FVB.129(B6)-Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J |
026431 | NOD.129(B6)-Gt(ROSA)26Sortm1(CAG-cas9*,-EGFP)Fezh/J |
026557 | NOD.129(Cg)-Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J |
037456 | NOD.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/YgchJ |
010675 | STOCK Gt(ROSA)26Sortm1(CAG-EGFP)Fsh/Mmjax |
010701 | STOCK Gt(ROSA)26Sortm1.1(CAG-EGFP)Fsh/Mmjax |
022386 | STOCK Gt(ROSA)26Sortm1.1(CAG-EGFP/Rpl10a,-birA)Wtp/J |
010812 | STOCK Gt(ROSA)26Sortm1.2(CAG-EGFP)Fsh/Mmjax |
018906 | STOCK Gt(ROSA)26Sortm3(CAG-flpo/ERT2)Alj/J |
007576 | STOCK Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J |
024107 | STOCK Gt(ROSA)26Sortm5(ACTB-tTA)Luo Igs7tm93.1(tetO-GCaMP6f)Hze/HzeJ |
024112 | STOCK Gt(ROSA)26Sortm5(ACTB-tTA)Luo Igs7tm94.1(tetO-GCaMP6s)Hze/J |
014552 | STOCK Nkx2-1tm1.1(cre/ERT2)Zjh/J |
39 strains use this protocol |