For in-depth product & services help, ask our
Technical Information Scientists
Mutant = 224 bp
Heterozygote = 165 bp and 224 bp
Wild type = 165 bp
This assay may be capable of distinguishing hemi from hom. Transgene insertion site is known to be on mouse Chr 2.
This assay is designed around this insertion site, but it has not been tested on hom animals.
This assay is NOT able to be used for transgene copy number evaluation. If this is required, it is suggested to type by qPCR.
>chr2:80896333-80896497 165bp GCCATGGTAGATAATGGCTTTC TTGGCTTACATATGGATTTTTGA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
37889 | GCC ATG GTA GAT AAT GGC TTT C | Common | A, B | mChr2 | ||
37890 | TTG GCT TAC ATA TGG ATT TTT GA | Wild type Reverse | A | mChr2 | ||
37891 | TTC TGA TGC AAA CCT CTT CC | Mutant Reverse | B | LRRK2 |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTPS-kapa | 0.26 mM |
37889 | 0.50 uM |
37890 | 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 |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTPS-kapa | 0.26 mM |
37889 | 0.50 uM |
37891 | 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 |