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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR0160 | TCT GGA CGA AGA GCA TCA GGG | Mutant Forward | A | |||
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 |
---|---|
002454 | B10.129P2(B6)-B2mtm1Unc/J |
003533 | B6.129P-B2mtm1Unc-rs2J/J |
002087 | B6.129P2-B2mtm1Unc/DcrJ |
002070 | B6;129P2-B2mtm1Unc/J |
002449 | BXSB.129P2(B6)-B2mtm1Unc/J |
002420 | C.129P2(B6)-B2mtm1Unc/J |
004040 | C3.129P2(B6)-B2mtm1Unc/Dcr |
002439 | C3.129P2(B6)-B2mtm1Unc/J |
002453 | MRL.129P2(B6)-B2mtm1Unc/J |
002455 | MRL.Cg-B2mtm1Unc Faslpr |
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 |
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 |
010636 | NOD.Cg-B2mtm1Unc Prkdcscid Il2rgtm1Wjl/SzJ |
018907 | NOD.Cg-B2mtm1Unc Tg(B2M)55Hpl Tg(HLA-B*0702)#Chmb/TdilJ |
003355 | NOD.Cg-B2mtm1Unc Tg(B2M)55Hpl/Dvs |
002570 | NOD.Cg-Prkdcscid B2mtm1Unc/J |
002452 | SJL.129P2(B6)-B2mtm1Unc/DcrJ |
21 strains use this protocol |