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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR8744 | CAA ATG TTG CTT GTC TGG TG | Internal Positive Control Forward | A | |||
oIMR8745 | GTC AGT CGA GTG CAC AGT TT | Internal Positive Control Reverse | A | |||
oIMR8746 | CGC TGT GGG GCA TTT TAC TTT AG | Transgene Forward | A | |||
oIMR8747 | CAT GTC CAG ATC GAA ATC GTC | Transgene Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
oIMR8744 | 0.50 uM |
oIMR8745 | 0.50 uM |
oIMR8746 | 0.50 uM |
oIMR8747 | 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 |
---|---|
016176 | B6(Cg)-Tg(tetO-Per2)2Jt/J |
028295 | B6.Cg-Tg(tetO-DRD3)3-10Kndl/J |
034533 | B6.FVB-Tg(EmuSR-tTa)83Bop/DwfJ |
017719 | C3HeB/FeJ-Tg(tetO-TAg)1Efr/J |
034802 | FVB/N-Tg(MMTV-tTA)25754Kuw/Mmjax |
004602 | NOD.Cg-Tg(Ins2-rtTA)2Doi/DoiJ |
005734 | NOD/Lt-Tg(Ins2-rtTA)1Ach/AchJ |
006999 | STOCK Dbttm1Geh Tg(Cebpb-tTA)5Bjd Tg(tetO-DBT)A1Geh/J |
018999 | STOCK Gt(ROSA)26Sortm1(tTA,tetO-Mir155)Fjsl/J |
024854 | STOCK Tg(Camk2a-tTA)1Mmay Fgf14Tg(tetO-MAPT*P301L)4510Kha/J |
003272 | STOCK Tg(Cebpb-tTA)5Bjd/J |
003273 | STOCK Tg(CMV-rtTA)4Bjd/J |
025672 | STOCK Tg(tetO-Fgfr2b/Igh)1.3Jaw/J |
13 strains use this protocol |