For in-depth product & services help, ask our
Technical Information Scientists
This assay will NOT distinguish hemizygous from homozygous transgenic animals.
Transgene = 170 bp
Internal positive control = 217 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
21221 | GAC AAA ATG GTG AAG GTC GG | Internal Positive Control Forward | A | |||
21222 | CAA AGG CGG AGT TAC CAG AG | Internal Positive Control Reverse | A | |||
29967 | GGT CTC CCA AAG GTA GAG TCG | Transgene Reverse | A | |||
oIMR9102 | ACA TGG TCC TGC TGG AGT TC | Transgene Forward | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
21221 | 0.50 uM |
21222 | 0.50 uM |
29967 | 0.50 uM |
oIMR9102 | 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 |
---|---|
029296 | FVB.Cg-Tg(tetO/Prnp-GLRA1*,-EYFP)9487Jjan/J |
029297 | FVB.Cg-Tg(tetO/Prnp-GLRA1*,-EYFP)9492Jjan/J |
029298 | FVB.Cg-Tg(tetO/Prnp-GLRA1*,-EYFP)9511Jjan/J |
029299 | FVB.Cg-Tg(tetO/Prnp-GLRA1*,-EYFP)9518Jjan/J |
029300 | FVB.Cg-Tg(tetO/Prnp-GLRA1*,-EYFP)9522Jjan/J |
029301 | FVB.Cg-Tg(tetO/Prnp-GLRA1*,-EYFP)9531Jjan/J |
029302 | FVB.Cg-Tg(tetO/Prnp-GLRA1*,-EYFP)9542Jjan/J |
7 strains use this protocol |