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Mutant = ~110 bp
Heterozygote = ~110 bp and 321 bp
Wild type = 321 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
23993 | TGG ATG GTG GTA TAC TCA GAG C | Common | A | |||
23994 | CAG CCT CTG TTC CAC ATA CAC T | Mutant Forward | A | |||
23995 | AGG CTT AGA GGT GCA AGC TG | Wild type Forward | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
23993 | 0.50 uM |
23994 | 0.50 uM |
23995 | 0.50 uM |
Glycerol | 6.50 % |
Dye | 1.00 X |
Kapa 2G HS taq polym | 0.03 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 65.0 | -- | -0.5 C per cycle decrease |
4 | 68.0 | -- | |
5 | -- | repeat steps 2-4 for 10 cycles (Touchdown) | |
6 | 94.0 | -- | |
7 | 60.0 | -- | |
8 | 72.0 | -- | |
9 | -- | repeat steps 6-8 for 28 cycles | |
10 | 72.0 | -- | |
11 | 10.0 | -- | hold |
Stock Number | Strain Name |
---|---|
002080 | 129-Trp53tm1Tyj/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 |
9 strains use this protocol |