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Mutant = 340 bp
Heterozygote = 340 bp and 650 bp
Wild type = 650 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR8052 | GCG AAG AGT TTG TCC TCA ACC | Mutant | A | SA site | ||
oIMR8545 | AAA GTC GCT CTG AGT TGT TAT | Common | A, B | ROSA26 | ||
oIMR8546 | GGA GCG GGA GAA ATG GAT ATG | Wild type | B | ROSA26 |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTPS-kapa | 0.26 mM |
oIMR8052 | 0.50 uM |
oIMR8545 | 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 |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTPS-kapa | 0.26 mM |
oIMR8545 | 0.50 uM |
oIMR8546 | 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 |
---|---|
009582 | B6(129S4)-Et(cre/ERT2)1645Rdav/J |
009585 | B6(129S4)-Et(cre/ERT2)2047Rdav/J |
012687 | B6(129S4)-Tg(SYN1-icre/mRFP1)9934Rdav/J |
012580 | B6.129P2(Cg)-Hprt1tm59(Ple123-lacZ)Ems/Mmjax |
012581 | B6.129P2(Cg)-Hprt1tm62(Ple153-lacZ)Ems/Mmjax |
012583 | B6.129P2(Cg)-Hprt1tm68(Ple127-lacZ)Ems/Mmjax |
013586 | B6.129P2-Gt(ROSA)26Sortm1Nik/J |
003474 | B6.129S4-Gt(ROSA)26Sortm1Sor/J |
007743 | B6.129S4-Gt(ROSA)26Sortm3(phiC31*)Sor/J |
032087 | B6.129S6(Cg)-Gt(ROSA)26Sortm1(DTA)Jpmb/J |
014602 | B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm1(tetO-mCherry)Eggn/J |
023749 | B6.Cg-Gt(ROSA)26Sortm1(rtTA*M2)Jae Tg(tetO-Pou5f1,-Sox2,-Klf4,-Myc)1Srn/J |
026294 | B6.Cg-Gt(ROSA)26Sortm2.2Ksvo/J |
010527 | B6;129-Gt(ROSA)26Sortm1(DTA)Mrc/J |
008883 | B6;129-Gt(ROSA)26Sortm1(SNCA*A53T)Djmo/TmdJ |
008889 | B6;129-Gt(ROSA)26Sortm2(SNCA*119)Djmo/TmdJ |
022516 | B6;129S-Gt(ROSA)26Sortm1(Cdkn1c)Jfpa/J |
003309 | B6;129S4-Gt(ROSA)26Sortm1Sor/J |
019101 | B6N.129S4(B6)-Gt(ROSA)26Sortm1Sor/CjDswJ |
019013 | B6N.129S6(Cg)-Gt(ROSA)26Sortm2(EGFP/cre)Alj/J |
007900 | C57BL/6-Gt(ROSA)26Sortm1(HBEGF)Awai/J |
011013 | STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm5(tetO-Pou5f1,-Klf4,-Myc)Jae/J |
018903 | STOCK Gt(ROSA)26Sortm2(EGFP/cre)Alj/J |
012584 | STOCK Hprt1tm69(Ple134-lacZ)Ems/Mmjax |
24 strains use this protocol |