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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR0180 | GCC TAG CCG AGG GAG AGC CG | Common | A | |||
oIMR0181 | TGT GAC TTG GGA GCT CTG CAG C | Wild type Reverse | A | |||
oIMR0182 | GCC GCC CCG ACT GCA TCT | Mutant Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
oIMR0180 | 0.50 uM |
oIMR0181 | 0.50 uM |
oIMR0182 | 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 |
---|---|
002246 | B6.129-Apoetm1Unc Ldlrtm1Her/J |
002052 | B6.129P2-Apoetm1Unc/J |
004631 | B6.Cg-Apoetm1Unc Cdh18Tg(GFAP-APOE_i4)1Hol/J |
007224 | B6.Cg-Apoetm1Unc (D12Mit182-D12Mit2)/Pgn |
008525 | B6.Cg-Tg(Cyp21a1-Apoe)619Fet/J |
004632 | B6.Cg-Tg(GFAP-APOE_i2)14Hol Apoetm1Unc/J |
004633 | B6.Cg-Tg(GFAP-APOE_i3)37Hol Apoetm1Unc/J |
002878 | B6;129-Apobtm1Sgy Apoetm1Unc/J |
002879 | B6;129-Apobtm2Sgy Apoetm1Unc/J |
002245 | B6;129-Apoetm1Unc Ldlrtm1Her/J |
004362 | B6;129-Scarb1tm1Kri Apoetm1Unc/J |
007067 | D2.129P2(B6)-Apoetm1Unc/J |
029818 | STOCK Apoetm1Unc Tg(APOE3*R224Q)1Her/J |
007069 | STOCK Apoetm1Unc/J |
14 strains use this protocol |