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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
>chr11:69589085+69589199 115bp GGTCACCTGTAGTGAGGTAGGG CCCATGCAGGAGCTATTACAC
Mut= 153 bp
Wt= 115 bp
Fam=Mut
Hex=Wt
Wt Sequence:
ggctgctgcaggtcacctgtagtgaggtagggagcgacttcacctggatcctgtgtcttcccccagGCCGGCTCTGAGTATACCACCATCCACTACAAGTACATGTGTAATAGCTCCTGCATGGGGGGCATG
Mutant Sequence:
TCATTCCTCCCACTCATGATCTATAGATCTATAGATCTCTCGTGGGATCATTGTTTTTCTCTTGATTCCCACTTTGTGGTTCTAAGTACTGTGGTTTCCAAATGTGTCAGTTTCATAGCCTGAAGAACGAGATCAGCAGCCTCTGTTCCACATACACTTCATTCTCAGTATTGTTTTGCCAAGTTCTAATTCCATCAGAAGCTGACTCTAGAGGATCCTGTGTCTTCCCCCAGGCCGGCTCTGAGTATACCACCATCCACTACAAGTACATGTGTAATAGCTCCTGCATGGGGGGCATGAACCGCCGACCTATCCTTACCATCATCACACTGGAAGACTCCAGGTAGGAAGGCGCGTGGTAGGTTAGGTTAGCCTGTTTCTTCCCCAGCTTCTGC
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
52606 | GGT CAC CTG TAG TGA GGT AGG G | Wild type Forward | A | |||
52607 | CCC ATG CAG GAG CTA TTA CAC A | Common | A | |||
52608 | CCT CTG TTC CAC ATA CAC TTC A | Mutant Forward | A | |||
52609 | Fluorophore-1 | AGC GAC TTC ACC TGG ATC C | Quencher-1 | WT Probe | ||
52610 | Fluorophore-2 | CCA TCA GAA GCT GAC TCT AGA GGA TCC | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
52606 | 0.40 uM |
52607 | 0.40 uM |
52608 | 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 |
---|---|
002080 | 129-Trp53tm1Tyj/J |
008651 | 129S-Trp53tm3Tyj/J |
002101 | B6.129S2-Trp53tm1Tyj/J |
027678 | B6;129-Trp53tm1Tyj Nf1tm1Tyj Suz12Gt(Betageo)1Khe/KcichJ |
008191 | B6;129S2-Trp53tm1Tyj Nf1tm1Tyj/J |
002103 | B6;129S2-Trp53tm1Tyj/J |
002526 | C.129S2(B6)-Trp53tm1Tyj/J |
002547 | C3Ou.129S2(B6)-Trp53tm1Tyj/J |
002899 | FVB.129S2(B6)-Trp53tm1Tyj/J |
017530 | STOCK Igs2tm2(ACTB-tdTomato,-EGFP)Luo Trp53tm1Tyj Nf1tm1Par/J |
10 strains use this protocol |