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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR0113 | CAT CAG CCC TAA TCC ATC TGA | Transgene Forward | A | SOD1 | ||
oIMR0114 | CGC GAC TAA CAA TCA AAG TGA | Transgene Reverse | A | SOD1 | ||
oIMR7338 | CTA GGC CAC AGA ATT GAA AGA TCT | Internal Positive Control Forward | A | |||
oIMR7339 | GTA GGT GGA AAT TCT AGC ATC ATC C | Internal Positive Control Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
oIMR0113 | 0.50 uM |
oIMR0114 | 0.50 uM |
oIMR7338 | 0.50 uM |
oIMR7339 | 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 |
---|---|
008342 | B6.Cg-Tg(SOD1*G37R)42Dpr/J |
008248 | B6.Cg-Tg(SOD1*G85R)148Dwc/J |
004435 | B6.Cg-Tg(SOD1*G93A)1Gur/J |
002297 | B6SJL-Tg(SOD1)2Gur/J |
002300 | B6SJL-Tg(SOD1*G93A)dl1Gur/J |
002726 | B6SJL-Tg(SOD1*G93A)1Gur/J |
002628 | C57BL/6-Tg(SOD1)10Cje/J |
7 strains use this protocol |