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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.
Mut-130bp
WT-94bp
Fam=Mut
Hex=wt
Cy5=IPC
Mutant sequence, including junction:
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
19475 | AAG AGA TTA CTT CTG GCT GTC AG | Common | A | |||
19476 | Fluorophore-1 | AAA GCG CCT CCC CTA CCC G | Quencher-1 | MUT Probe | PGK | |
19477 | CTC TGG GGT TTC TGG GG | Wild type Reverse | A | |||
19478 | Fluorophore-2 | TGG AGC TGG ACA ACA AAT GTC TGG T | Quencher-2 | WT Probe | ||
oIMR3914 | ATG CTC CAG ACT GCC TTG | Mutant Reverse | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
19475 | 0.40 uM |
19477 | 0.40 uM |
oIMR3914 | 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 |
---|---|
032088 | B6.129S-Rag1tm1Mom Cd47tm1Fpl Il2rgtm1Wjl/SzJ |
025730 | B6.129S-Rag2tm1Fwa Cd47tm1Fpl Il2rgtm1Wjl/J |
003174 | B6.129S4-Il2rgtm1Wjl/J |
039001 | B6.Cg-Rag2tm1Fwa Sgshmps3a Cd47tm1Fpl Il2rgtm1Wjl/JnolJ |
003169 | C.129S4-Il2rgtm1Wjl/J |
030547 | NOD.Cg-B2mtm1Unc Prkdcscid H2dlAb1-Ea Il2rgtm1Wjl/SzJ |
010636 | NOD.Cg-B2mtm1Unc Prkdcscid Il2rgtm1Wjl/SzJ |
038279 | NOD.Cg-Capn3em5Lutzy Prkdcscid Il2rgtm1Wjl/VitaJ |
031900 | NOD.Cg-Cmahem1Guru Prkdcscid Il2rgtm1Wjl/GuruJ |
028615 | NOD.Cg-Fcgrttm1Dcr Prkdcscid Il2rgtm1Wjl Tg(FCGRT)32Dcr/J |
035844 | NOD.Cg-Flt3em1Akp Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(HLA-A/H2-D/B2M)1Dvs/J |
035843 | NOD.Cg-Flt3em1Akp Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J |
035842 | NOD.Cg-Flt3em1Akp Prkdcscid Il2rgtm1Wjl/J |
037322 | NOD.Cg-Flt3em2Mvw Prkdcscid Il2rgtm1Wjl Tg(FLT3LG)7Sz Tg(IL15)1Sz/J |
033367 | NOD.Cg-Flt3em2Mvw Prkdcscid Il2rgtm1Wjl Tg(FLT3LG)7Sz/SzJ |
026263 | NOD.Cg-Foxn1em1Dvs Prkdcscid Il2rgtm1Wjl/J |
029819 | NOD.Cg-Gcgem1Dvs Prkdcscid Il2rgtm1Wjl/DvsJ |
035959 | NOD.Cg-Gt(ROSA)26Sorem27(KRT18-ACE2)Mvw Prkdcscid Il2rgtm1Wjl/MvwJ |
038123 | NOD.Cg-Gt(ROSA)26Sorem28(KRT18-ACE2)Mvw Prkdcscid Il2rgtm1Wjl/MvwJ |
036151 | NOD.Cg-Gt(ROSA)26Sorem5Mvw Prkdcscid Il2rgtm1Wjl/MvwJ |
030511 | NOD.Cg-Hc1 Prkdcscid Il2rgtm1Wjl/SzJ |
014553 | NOD.Cg-Hgftm1.1(HGF)Aveo Prkdcscid Il2rgtm1Wjl/J |
035868 | NOD.Cg-Iduaem1Osp Prkdcscid Il2rgtm1Wjl/J |
035007 | NOD.Cg-Iduatm1Clk Prkdcscid Il2rgtm1Wjl/J |
026622 | NOD.Cg-KitW-41J Tyr + Prkdcscid Il2rgtm1Wjl/ThomJ |
026497 | NOD.Cg-KitW-41J Prkdcscid Il2rgtm1Wjl/WaskJ |
009617 | NOD.Cg-Mcph1Tg(HLA-A2.1)1Enge Prkdcscid Il2rgtm1Wjl/SzJ |
032474 | NOD.Cg-Prkdcscid Abcd1em2Lutzy Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ |
027905 | NOD.Cg-Prkdcscid Cd274tm1Shr Il2rgtm1Wjl/J |
030610 | NOD.Cg-Prkdcscid Cybbem1Hmal Il2rgtm1Wjl/HmalJ |
031566 | NOD.Cg-Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl Tg(H2-Ea-HLA-DRB1*0401*)1Dv/SzJ |
026936 | NOD.Cg-Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv Tg(INS*)172Dvs/DvsJ |
026561 | NOD.Cg-Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv/SzJ |
021885 | NOD.Cg-Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl/SzJ |
025216 | NOD.Cg-Prkdcscid H2-K1b-tm1Bpe H2-Ab1g7-em1Mvw H2-D1b-tm1Bpe Il2rgtm1Wjl/SzJ |
023848 | NOD.Cg-Prkdcscid H2-K1b-tm1Bpe H2-D1b-tm1Bpe Il2rgtm1Wjl/SzJ |
039055 | NOD.Cg-Prkdcscid H2-K1d-em1Dvs H2-Ab1g7-em1Dvs H2-D1b-em5Dvs Il2rgtm1Wjl Tg(HLA-DQA1,HLA-DQB1)1Dv/DvsJ |
032927 | NOD.Cg-Prkdcscid H2-K1d-em1Dvs H2-Ab1g7-em1Dvs H2-D1b-em5Dvs Il2rgtm1Wjl/SzJ |
026222 | NOD.Cg-Prkdcscid Hprt1em1Mvw Il2rgtm1Wjl/MvwJ |
028657 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(CSF1)3Sz/J |
014570 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(HLA-A/H2-D/B2M)1Dvs/SzJ |
030890 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(IL15)1Sz/SzJ |
035970 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(YAC128)53Hay/JnolJ |
035424 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Ace2em1Lutzy/J |
035002 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Ace2tm1(ACE2)Dwnt/J |
017637 | NOD.Cg-Prkdcscid Il2rgtm1Wjl H2-Ab1b-tm1Doi Tg(HLA-DRB1)31Dmz/SzJ |
012480 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Hprt1b-m3/EshJ |
034859 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CAG-cre/Esr1*)5Amc/J |
021937 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CAG-EGFP)1Osb/SzJ |
013062 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ |
028654 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CSF1)3Sz/SzJ |
029295 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(H2-Ea-HLA-DRB1*0401*)1Dv/SzJ |
028655 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(IL6)1Sz/SzJ |
027976 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(Ins2-HBEGF)6832Ugfm/SzJ |
017830 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(PGK1-KITLG*220)441Daw/SzJ |
028842 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(SERPINA1*E342K)#Slcw/SzJ |
005557 | NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ |
034569 | NOD.Cg-Prkdcscid Otcem1Bshep Il2rgtm1Wjl/J |
011066 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CSF2)2Ygy Tg(IL3)1Ygy Tg(KITLG)3Ygy/YgyJGckRolyJ |
036917 | NOD.Cg-Prltm1.1(PRL)Rui Prkdcscid Il2rgtm1Wjl/J |
030442 | NOD.Cg-Rag1tm1Mom Dmdmdx-4Cv Il2rgtm1Wjl/McalJ |
018454 | NOD.Cg-Rag1tm1Mom Fahem1Mvw Il2rgtm1Wjl/MvwJ |
026062 | NOD.Cg-Rag1tm1Mom Fancaem1Dvs Il2rgtm1Wjl/SzJ |
033127 | NOD.Cg-Rag1tm1Mom Flt3tm1Irl Mcph1Tg(HLA-A2.1)1Enge Il2rgtm1Wjl/J |
035855 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(HLA-A/H2-D/B2M)Dvs Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito/J |
030533 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(SLC10A1)15Mvw/MvwJ |
026014 | NOD.Cg-Rag1tm1Mom KitW-41J Il2rgtm1Wjl/EavJ |
030443 | NOD.Cg-Rag1tm1Mom Sgcatm1Kcam Il2rgtm1Wjl/McalJ |
029663 | NOD.Cg-Rag1tm1Mom Dysfprmd Il2rgtm1Wjl/McalJ |
024099 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J |
017914 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito/ScasJ |
007799 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ |
014568 | NOD.Cg-Rag1tm1Mom Ins2Akita Il2rgtm1Wjl/SzJ |
037275 | NOD.Cg-Rag2em8Lutzy H2-K1b-tm1Bpe H2-Ab1g7-em1Mvw H2-D1b-tm1Bpe Il2rgtm1Wjl/J |
037276 | NOD.Cg-Rag2em8Lutzy Il2rgtm1Wjl/J |
033704 | NOD.Cg-Tlr4lps-del Prkdcscid Il2rgtm1Wjl/SzJ |
028616 | NOD.Cg-Tg(CAG-FCGRT)276Dcr Fcgrttm1Dcr Prkdcscid Il2rgtm1Wjl/J |
026997 | NOD.Cg-Tg(CD68-EGFP)1Drg Prkdcscid Il2rgtm1Wjl/J |
030331 | NOD.Cg-Tg(HLA-DRA*0101,HLA-DRB1*0101)1Dmz Prkdcscid H2-Ab1b-tm1Doi Il2rgtm1Wjl/SzJ |
012479 | NOD.Cg-Tg(HLA-DRA*0101,HLA-DRB1*0101)1Dmz Prkdcscid Il2rgtm1Wjl/GckRolyJ |
034901 | NOD.Cg-Tg(K18-ACE2)2Prlmn Prkdcscid Il2rgtm1Wjl/J |
81 strains use this protocol |