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>chr11:104265226+104265523 298bp TTCTTAACCGCTGAGCCATC CCTTTGGGATTTTAACCATGTG
Mutant = 151 bp
Heterozygote = 151 bp and 298 bp
Wild type = 298 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
49773 | TTC TTA ACC GCT GAG CCA TC | Wild type Forward | A | Mapt F | ||
49774 | CCT TTG GGA TTT TAA CCA TGT G | Wild type Reverse | A | Mapt R | ||
49775 | TCA CTT ACA CGA GGC ATC CA | Mutant Forward | A | MAPT F | ||
49776 | AAC TGA CCA CGG TTC TCC AC | Mutant Reverse | A | MAPT R |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
49773 | 0.50 uM |
49774 | 0.50 uM |
49775 | 0.50 uM |
49776 | 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 |