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Mutant Tm = 88.7°C +/- 1.0°C
Wild type Tm = 83.1°C+/- 1.0°C
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR0160 | TCT GGA CGA AGA GCA TCA GGG | Mutant Forward | A | neo | ||
oIMR0184 | TAT CAG TCT CAG TGG GGG TG | Common | A | |||
oIMR0185 | CTG AGC TCT GTT TTC GTC TG | Wild type Forward | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
oIMR0160 | 0.50 uM |
oIMR0184 | 0.50 uM |
oIMR0185 | 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 |
---|---|
002087 | B6.129P2-B2mtm1Unc/DcrJ |
002420 | C.129P2(B6)-B2mtm1Unc/J |
005356 | NOD.129(B6)-B2mtm1Unc Ciitatm1Ccum/BhsJ |
010565 | NOD.129P2(B6)-B2mtm1Unc Tg(HLA-A24/H2-D/B2M)3Dvs/J |
002309 | NOD.129P2(B6)-B2mtm1Unc/J |
027849 | NOD.Cg-B2mtm1Unc Tg(B2M)55Hpl Tg(HLA-A*1101/H2-Kb)#Sette/TdilJ |
008542 | NOD.Cg-B2mtm1Unc Tg(GFAP-B2m)9Mdos/J |
006611 | NOD.Cg-B2mtm1Unc Tg(HLA-A/H2-D/B2M)1Dvs/DvsJ |
004548 | NOD.Cg-B2mtm1Unc Mcph1Tg(HLA-A2.1)1Enge Tg(B2M)55Hpl/DvsJ |
030547 | NOD.Cg-B2mtm1Unc Prkdcscid H2dlAb1-Ea Il2rgtm1Wjl/SzJ |
010636 | NOD.Cg-B2mtm1Unc Prkdcscid Il2rgtm1Wjl/SzJ |
018907 | NOD.Cg-B2mtm1Unc Tg(B2M)55Hpl Tg(HLA-B*0702)#Chmb/TdilJ |
002570 | NOD.Cg-Prkdcscid B2mtm1Unc/J |
13 strains use this protocol |