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HET = 268 bp and 763 or 162 bp
MUT A = 763 bp and 162 bp in agouti (A) allele
WT nonagouti (a) = 268 bp
>chr2:155017745+155018012 268bp AGTCCCACAGAGACGAGACC GAGGGGACTATAATTCAGCTTTTAC
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
50528 | AGT CCC ACA GAG ACG AGA CC | Wild type Forward | A | |||
50529 | GAG GGG ACT ATA ATT CAG CTT TTA C | Wild type Reverse | A | |||
oIMR7876 | GGC CAC ACT TAG GGA GTT CA | Mutant Forward | B | |||
oIMR7932 | GGT TGG CCA CCA TGT CTA GT | Mutant Reverse | B |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTPS-kapa | 0.26 mM |
50528 | 0.50 uM |
50529 | 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 |
oIMR7876 | 0.50 uM |
oIMR7932 | 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 |
---|---|
000202 | AEJ/Gn-bd/J |
000199 | AEJ/GnLeJ |
000427 | B10.CE-H13b Aw/(30NX)SnJ |
000420 | B10.LP-H13b Aw/Sn |
000593 | B6 x B6CBCa Aw-J/A-Grid2Lc T(2;6)7Ca MitfMi-wh/J |
000502 | B6 x B6CBCa Aw-J/A-Myo5aflr Gnb5flr/J |
000599 | B6 x B6CBCa Aw-J/A-T(5;13)264Ca KitW-v/J |
002016 | B6(Cg)-Aw-J EdaTa-6J Chr YB6-Sxr/EiJ |
000600 | B6-Gpi1b x B6CBCa Aw-J/A-T(7;15)9H Gpi1a/J |
001809 | B6.Cg-Aw-J EdaTa-6J +/+ ArTfm/J |
000552 | B6.Cg-Aw-J EdaTa-6J Sxr |
001730 | B6.Cg-Aw-J EdaTa-6J Sxrb Hya-/J |
000841 | B6.Cg-Aw-J EdaTa-By/J |
005505 | B6.Cg-Ay Slc7a11sut/LmLlp |
000021 | B6.Cg-Ay/J |
004200 | B6;CBACa Aw-J/A-Npr2cn-2J/GrsrJ |
100409 | B6129PF1/J-Aw-J/Aw |
000505 | B6C3 Aw-J/A-Bloc1s5mu/J |
000314 | B6CBACa Aw-J/A-EdaTa/J-XO |
000288 | B6CBACa Aw-J/A-Tgm3we a Mafbkr/J |
000501 | B6CBACa Aw-J/A-Aifm1Hq/J |
001046 | B6CBACa Aw-J/A-Grid2Lc/J |
000500 | B6CBACa Aw-J/A-Gs/J |
002703 | B6CBACa Aw-J/A-Hydinhy3/J |
000247 | B6CBACa Aw-J/A-Kcnj6wv/J |
000287 | B6CBACa Aw-J/A-Plp1jp EdaTa/J |
000515 | B6CBACa Aw-J/A-SfnEr/J |
000242 | B6CBACa Aw-J/A-spc/J |
001201 | B6CBACaF1/J-Aw-J/A |
002343 | B6EiC3Sn a/A-Otcspf/J |
000638 | C3FeB6 A/Aw-J-Sptbn4qv-J/J |
000200 | C3FeB6 A/Aw-J-Ankank/J |
001203 | C3FeB6F1/J A/Aw-J |
000664 | C57BL/6J |
000338 | C57BL/6J Aw-J-EdaTa-6J/J |
000569 | C57BL/6J-Aw-J-EdaTa +/+ ArTfm/J |
000051 | C57BL/6J-Aw-J/J |
002468 | KK.Cg-Ay/J |
000064 | STOCK a Tyrp1b Pmelsi/J |
001432 | STOCK a/a Tyrp1b Ndc1sks/Tyrp1b +/J |
001427 | STOCK Aw us/J |
001752 | STOCK T(7;15)9H/J |
42 strains use this protocol |