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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR7376 | CCA CAC GCG TCA CCT TAA TA | Mutant Reverse | A | |||
oIMR9573 | CTT GCA CTA CCA AAG CCA CA | Common | A | |||
oIMR9574 | GTT ATT GTC TTC CCG GCT GT | Wild type Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
oIMR7376 | 0.50 uM |
oIMR9573 | 0.50 uM |
oIMR9574 | 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 |
---|---|
004368 | 129(B6)-Il10tm1Cgn/J |
002250 | B10.129P2(B6)-Il10tm1Cgn/J |
002251 | B6.129P2-Il10tm1Cgn/J |
005912 | B6.Cg-Il10tm1Cgn (D3Mit49-D3Mit348)/Lt |
004333 | C.129P2(B6)-Il10tm1Cgn/J |
003968 | C3Bir.129P2(B6)-Il10tm1Cgn/LtJ |
036145 | CBA.129P2(B6)-Il10tm1Cgn/TrnrJ |
004291 | NOD.129P2(Cg)-Il10tm1Cgn Il4tm1Cgn/DvsJ |
004266 | NOD.129P2(Cg)-Il10tm1Cgn/DvsJ |
005346 | NOD.Cg-Il10tm1Cgn Casp1tm1Sesh Casp4del/LtJ |
004228 | NOD.Cg-Il10tm1Cgn Il4tm1Cgn/Dvs |
11 strains use this protocol |