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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
9767 | GAA ATG TCC GTT CGG TTG GCA GAA GC | Transgene Forward | A | Luc | ||
9768 | CCA AAA CCG TGA TGG AAT GGA ACA ACA | Transgene Reverse | A | Luc | ||
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 |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
9767 | 0.50 uM |
9768 | 0.50 uM |
oIMR8744 | 0.50 uM |
oIMR8745 | 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 |
---|---|
006100 | B10.Cg-H2k Tg(NFkB/Fos-luc)26Rinc/J |
033605 | B6.Cg-S1pr1tm4.1Rlp Gt(ROSA)26Sortm2.1(luc*/Arrb2*)Rlp Tyrc-2J/J |
023429 | B6.Cg-Tyrc-2J Cdkn1atm1Hpw/J |
006098 | B6.Cg-Tg(Il2/NFAT-luc)83Rinc/J |
006101 | B6.Cg-Tg(TRE/Prl-luc)31FlvRinc/J |
002709 | B6;C3-Tg(TettTALuc)1Dgs/J |
027662 | C57BL/6-Tyrc-Brd Tg(Gh1-luc/EGFP)D8Mrln/J |
007800 | FVB/N-Tg(Ins1-luc)VUPwrs/J |
032442 | FVB/N-Tg(tetO/CMV-Dek,-luc)317Siwe/J |
012370 | FVB/NJ-Tg(Hspa1a-luc,-EGFP)2Chco/J |
10 strains use this protocol |