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AAGCAGACACAGGAGAGAGGATGTTCTGTaaaacaagcatgtga
Aagga(tg/--)cg(atg/---)cgtgatgaaggattctttgctaaaGACACA
ATTATGGATCTGCAA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
20197 | AGT CGG TTG GAT GGT GTT GT | Forward | A | |||
20198 | AAT GGG CCT CTC TTT CCA GT | Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTPS-kapa | 0.26 mM |
20197 | 0.50 uM |
20198 | 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 |
---|---|
002845 | C.B-H2b Tg(H2-Dd)D8Gja/LilJ |
001952 | C.B10-H2b/LilMcdJ |
000438 | C3.SW-H2b/SnJ |
000666 | C57BL/10SnJ |
004353 | C57BL/6-Tg(UBC-GFP)30Scha/J |
000664 | C57BL/6J |
005304 | C57BL/6NJ |
007076 | CByJ.B6-Tg(UBC-GFP)30Scha H2b/J |
002591 | NOD.B10Sn-H2b/J |
9 strains use this protocol |