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>chr11:104188843+104189134 292bp GCTCATCCCCAATTTCTTGA GTTGTTGCCTGTCCCAGACT
Mutant = 163 bp
Heterozygote = 163 bp and 292 bp
Wild type = 292 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
49779 | GCT CAT CCC CAA TTT CTT GA | Wild type Forward | A | Wt F | ||
49780 | GTT GTT GCC TGT CCC AGA CT | Wild type Reverse | A | Wt R | ||
49781 | CGC AGA TGT TGC AGT AGG AA | Mutant Forward | A | Mut F | ||
49782 | TCG GAG CTG AAC AGG ATA CA | Mutant Reverse | A | Mut R |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
49779 | 0.50 uM |
49780 | 0.50 uM |
49781 | 0.50 uM |
49782 | 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 |
---|---|
036664 | B6(Cg)-Tc(HSA17*)1Mdk/J |
038908 | B6.Cg-Tc(HSA17)2Mdk Appem1Adiuj/J |
039175 | B6J.B6N(CBA)-Tc(HSA17)1Mdk Psen1tm1.1(PSEN1)Mdk Apptm1.1(APP)Mdk/J |
033668 | B6J.B6N(CBA)-Tc(HSA17)1Mdk/J |
035794 | B6J.B6N(Cg)-Tc(HSA17*N279K)1Mdk/J |
035398 | B6J.B6N-Tc(HSA17)2Mdk/J |
037420 | B6J.B6N-Tc(HSA17*P301L)1Mdk/J |
7 strains use this protocol |