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This assay cannot distinguish hemi from hom
Tg= 124 bp
IPC = 74 bp
Tg Sequence:
TGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGTTATTGTGCTGTCTCATCATTTTGGCAAAGAATTCGGGCCCCCCCTCGAGGGGCAGAGCCGATCCTGTACACTTTACTTAAAACCATTATCTGAGTGTGAAATGTTCAATTTACTGACCGTACACCAAAATTTGCCTAG
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
34918 | TGT GCT GTC TCA TCA TTT TGG | Transgene Forward | A | |||
52581 | TTG GTG TAC GGT CAG TAA ATT G | Transgene Reverse | A | |||
52582 | Fluorophore-1 | AGG GGC AGA GCC GAT CCT | Quencher-1 | Tg Probe | ||
oIMR1544 | CAC GTG GGC TCC AGC ATT | Internal Positive Control Forward | A | |||
oIMR3580 | TCA CCA GTC ATT TCT GCC TTT G | Internal Positive Control Reverse | A | |||
TmoIMR0105 | Fluorophore-2 | CCA ATG GTC GGG CAC TGC TCA A | Quencher-2 | IC Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
34918 | 0.40 uM |
52581 | 0.40 uM |
oIMR1544 | 0.40 uM |
oIMR3580 | 0.40 uM |
Wt Probe | 0.15 uM |
Mutant Probe | 0.15 uM |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 95.0 | -- | |
2 | 95.0 | -- | |
3 | 60.0 | -- | |
4 | -- | repeat steps 2-3 for 40 cycles | |
5 | 40.0 | -- | Forever |
Stock Number | Strain Name |
---|---|
007966 | B6.Cg-Smn1tm3(SMN2/Smn1)Mrph/J |
004682 | B6.Cg-Tg(CAG-cre/Esr1*)5Amc/J |
034156 | B6N(Cg)-Pex16tm1d(EUCOMM)Hmgu/LutzyJ |
019102 | B6N.Cg-Tg(CAG-cre/Esr1*)5Amc/CjDswJ |
017595 | FVB.Cg-Tg(CAG-cre/Esr1*)5Amc/J |
034859 | NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CAG-cre/Esr1*)5Amc/J |
034795 | NOD.Cg-Tg(CAG-cre/Esr1*)5Amc/J |
008783 | STOCK Grm7Tg(SMN2)89Ahmb Smn1tm3(SMN2/Smn1)Mrph Tg(CAG-cre/Esr1*)5Amc Tg(SMN2*delta7)4299Ahmb/J |
008813 | STOCK Trpa1tm2Kykw Tg(CAG-cre/Esr1*)5Amc/J |
004453 | STOCK Tg(CAG-cre/Esr1*)5Amc/J |
10 strains use this protocol |