For in-depth product & services help, ask our
Technical Information Scientists
Mutant = 257 bp
Heterozygote = 160 bp and 257 bp
Wild type = 160 bp
Two protocols are provided because of different band sizes.
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
28995 | CTG GTT GCC CCA GCT TC | Wild type Forward | A | |||
28996 | CGA TTT ATA TTC CTC CAG GGT A | Common | A | |||
28997 | GAG GAC ATT CCA ATC ATA GGC | Mutant Forward | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
28995 | 0.50 uM |
28996 | 0.50 uM |
28997 | 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 |
---|---|
019995 | B6.129P2-H2-K1b-tm1Bpe H2-D1b-tm1Bpe/DcrJ |
025216 | NOD.Cg-Prkdcscid H2-K1b-tm1Bpe H2-Ab1g7-em1Mvw H2-D1b-tm1Bpe Il2rgtm1Wjl/SzJ |
023848 | NOD.Cg-Prkdcscid H2-K1b-tm1Bpe H2-D1b-tm1Bpe Il2rgtm1Wjl/SzJ |
037275 | NOD.Cg-Rag2em8Lutzy H2-K1b-tm1Bpe H2-Ab1g7-em1Mvw H2-D1b-tm1Bpe Il2rgtm1Wjl/J |
4 strains use this protocol |