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Mutant = 226 bp
Heterozygote = 218 bp and 226 bp
Wild type = 218 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
19125 | CTT CCC TCT ACC CTG CCT TC | Wild type Forward | A | |||
19126 | GGA GAA CAG TGG ACA CAG TAA CA | Common | A | |||
oIMR8444 | GCC TGA AGA ACG AGA TCA GC | Mutant Forward | A | pPNT polyA |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
19125 | 0.50 uM |
19126 | 0.50 uM |
oIMR8444 | 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 |
---|---|
016842 | B6.129(Cg)-Fxntm1Mkn/J |
012329 | B6.Cg-Fxntm1.1Pand Fxntm1Mkn/J |
014162 | B6.Cg-Fxntm1Mkn Fxntm1Pand/J |
012910 | B6.Cg-Fxntm1Mkn Tg(FXN)YG22Pook/J |
012253 | B6.Cg-Fxntm1Mkn Tg(FXN)YG8Pook/J |
029721 | B6.Cg-Pvalbtm1(cre)Arbr Fxnem2Lutzy Fxnem2.1Lutzy/J |
008586 | B6.Cg-Tg(FXN*)1Sars Fxntm1Mkn/J |
031007 | STOCK Fxnem2.1Lutzy Tg(FXN)Y47Pook/J |
024097 | STOCK Fxntm1Mkn Tg(FXN)Y47Pook/J |
024113 | STOCK Fxntm1Mkn Tg(FXN)YG8Pook/2J |
010963 | STOCK Fxntm1Mkn Tg(FXN)YG22Pook/J |
008398 | STOCK Fxntm1Mkn Tg(FXN)YG8Pook/J |
030324 | STOCK Tg(FXN)YG8Pook/J |
018299 | STOCK Tg(FXN*)1Sars Fxntm1Mkn/J |
14 strains use this protocol |