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Mutant = ~179 bp
Heterozygote = ~179 bp and 351 bp
Wild type = 351 bp
Mut is GC rich
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
19799 | TAT GCA TCC CCA GTC TTT GG | Common | A | |||
19800 | CTA CCC AAC CAG CCC CTT AC | Wild type Reverse | A | |||
oIMR7770 | ATA GAT TCG CCC TTG TGT CC | Mutant Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
19799 | 0.50 uM |
19800 | 0.50 uM |
oIMR7770 | 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 |
---|---|
007070 | AK.129S7(B6)-Ldlrtm1Her/J |
006952 | B6.129-Akt2tm1.1Mbb Ldlrtm1Her/J |
002246 | B6.129-Apoetm1Unc Ldlrtm1Her/J |
006883 | B6.129S7-Ldlrtm1Her Sod2tm1Leb/J |
002207 | B6.129S7-Ldlrtm1Her/J |
006580 | B6.Cg-Ins2Akita Ldlrtm1Her/J |
022488 | B6.Cg-Ldlrtm1Her Tg(Tagln-Pth1r*)2Towl/J |
006877 | B6.Cg-Ldlrtm1Her Tg(H2-K-AKR1B1)1Tj/J |
006906 | B6.Cg-Lepob Ldlrtm1Her/J |
002245 | B6;129-Apoetm1Unc Ldlrtm1Her/J |
003000 | B6;129S-Ldlrtm1Her Apobtm2Sgy/J |
002465 | B6;129S-Lrpap1tm1Her Ldlrtm1Her/J |
002077 | B6;129S7-Ldlrtm1Her/J |
012845 | CByJ.129S7(B6)-Ldlrtm1Her/J |
007068 | D2.129S7(B6)-Ldlrtm1Her/J |
004192 | STOCK Mttptm2Sgy Ldlrtm1Her Apobtm2Sgy Tg(Mx1-cre)1Cgn/J |
16 strains use this protocol |