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Tg=241bp
Internal control = 314bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
9801 | CCT CCT GTT AGC TGG GCT TT | Transgene Forward | A | |||
9802 | ACC ACT CCC TCC TTG GTT TT | Transgene Reverse | A | |||
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 |
9801 | 0.50 uM |
9802 | 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 |
---|---|
024338 | B6.Cg-Sncatm1Rosl Tg(SNCA*A30P)192Rwm/J |
023837 | B6.Cg-Tg(SNCA)OVX37Rwm Sncatm1Rosl/J |
028808 | FVB(B6)-Tg(SNCA*)BXwy/J |
010710 | FVB;129S6-Sncatm1Nbm Tg(SNCA)1Nbm/J |
010788 | FVB;129S6-Sncatm1Nbm Tg(SNCA*A30P)1Nbm Tg(SNCA*A30P)2Nbm/J |
010799 | FVB;129S6-Tg(SNCA*A53T)1Nbm Sncatm1Nbm Tg(SNCA*A53T)2Nbm/J |
027227 | STOCK Gba1tm1Rlp Tg(SNCA*A53T)1Nbm Sncatm1Nbm Tg(SNCA*A53T)2Nbm/Mmjax |
7 strains use this protocol |