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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
>chr6:111099925+111100076 152bp CAAACCAAAGCTCCAGAATC GTGGCCAAATGTCAGTCACC
Mut= 158 bp
Wt= 152 bp
Fam=Mut
Hex=Wt
Wt Sequence:
tctacattactgacctaccagggatgaggcattaggggaaatttttccaactcagtgtcatagcttctgtgtgctgtatgaactgataagttaacttttaacactgtataaattctgaaaacaaaccaaagctccagaatctttgaaaaatgagtttttgtttaagaaaccagaaatatgtagcctcccaaattgtcTGtaccctctgagttctcaattatttttaagtgtctgtgctacttagaggtggccaggtgactgacatttggccactatgaaggaagaaac
Mutant Sequence:
GTATAAATTCTGAAAACAAACCAAAGCTCCAGAATCTTTGAAAAATGAGTTTTTGTTTAAGAAACCAGAAATATGTAGCCTCCCAAATTgtctCAAAGTAATCTCTGTCTGATCCCTTAATGAAGGATCTCTTCAAACACAGCAACTTTTGTCTGTCTTCTGAACGTAGGTACAGCTGTGGCTG
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
52364 | CAA ACC AAA GCT CCA GAA TC | Common | A | |||
52365 | GTG GCC AAA TGT CAG TCA CC | Wild type Reverse | A | |||
52366 | TGT ACC TAC GTT CAG AAG ACA GAC | Mutant Reverse | A | |||
52367 | Fluorophore-1 | CCC AAA TTG TCT GTA CCC TCT GAG | Quencher-1 | WT Probe | ||
52368 | Fluorophore-2 | CCC TTA ATG AAG GAT CTC TTC AAA CAC | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
52364 | 0.40 uM |
52365 | 0.40 uM |
52366 | 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 |
---|---|
007222 | B6.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN1*A2G)2023Ahmb/J |
006964 | B6.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2*delta7)4299Ahmb/J |
006773 | B6.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd/J |
006385 | B6;129P2-Syt1tm3Sud/J |
006386 | B6;129P2-Syt1tm5Sud/J |
008209 | FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(ACTA1-SMN)69Ahmb/J |
007968 | FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN1*A2G)2023Ahmb/2J |
005026 | FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN1*A2G)2023Ahmb/J |
007952 | FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2*delta7)4299Ahmb/2J |
005025 | FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2*delta7)4299Ahmb/J |
031906 | FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2-SMN1*T274I)5Ahmb/J |
007949 | FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd/2J |
005024 | FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd/J |
030214 | FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(Prnp-PLS3)14Ahmb Tg(SMN2*delta7)4299Ahmb/J |
030970 | FVB.Cg-Mapk10tm1Flv Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2*delta7)4299Ahmb/LganJ |
031907 | STOCK Grm7Tg(SMN2)89Ahmb Tg(SMN2-SMN1*Q282A)2Ahmb Smn1tm1Msd/J |
008203 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(ACTA1-SMN)63Ahmb/J |
032003 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(Hlxb9-GFP)1Tmj Tg(SMN2*delta7)4299Ahmb/J |
006570 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(Hlxb9-GFP)1Tmj/J |
008212 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(Prnp-SMN)92Ahmb/J |
025102 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(Rnu7-SMN2*,PGK1-EGFP)4Dasch/DaschMmjax |
018916 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN1-SMN2*)16Cll/CllJ |
031908 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2-SMN1*D44V)10Ahmb/J |
008783 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm3(SMN2/Smn1)Mrph Tg(CAG-cre/Esr1*)5Amc Tg(SMN2*delta7)4299Ahmb/J |
007951 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm3(SMN2/Smn1)Mrph Tg(SMN2*delta7)4299Ahmb/J |
007022 | STOCK Mnx1tm4(cre)Tmj Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2*delta7)4299Ahmb/J |
017600 | STOCK Tg(tetO-SMN2,-luc)#bAhmb/J |
27 strains use this protocol |