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This assay will NOT distinguish hemizygous from homozygous transgenic animals.
Transgene = ~300 bp
Internal positive control = 168 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
21157 | CTC CCA ACC CCA GAG GTA GT | Internal Positive Control Forward | A | |||
21159 | CTA TAG GGC CTG GGT CAG TG | Internal Positive Control Reverse | A | |||
oIMR1201 | GTG AAG GAA CCT TAC TTC TGT GGT G | Transgene Forward | A | |||
oIMR1202 | GTC CTT GGG GTC TTC TAC CTT TCT C | Transgene Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
21157 | 0.25 uM |
21159 | 0.25 uM |
oIMR1201 | 0.50 uM |
oIMR1202 | 0.50 uM |
Glycerol | 6.50 % |
Dye | 1.00 X |
Kapa 2G HS taq polym | 0.03 U/ul |
DNA |