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This assay cannot distinguish hemi from hom
This assay can distinguish GCaMP6f from GCaMP6s and GCaMP3
Tg= 95 bp
IPC = 74 bp
tcgcagaatttaaagaggAAttctccctatttgacaaggacggggatggga
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
31035 | GGA CCA ACT GAC TGA AGA GCA | Forward | A | |||
31036 | CCA GCT CCT TGG TTG TTA TTG | Reverse | A | |||
31038 | Fluorophore-1 | CAG AAT TTA AAG AGG AAT TCT CCC TAT | Quencher-1 | MUT 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 | |
31035 | 0.40 uM |
31036 | 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 |
---|---|
024103 | B6;129S6-Igs7tm93.1(tetO-GCaMP6f)Hze/J |
024339 | C57BL/6J-Tg(Thy1-GCaMP6f)GP5.11Dkim/J |
028281 | C57BL/6J-Tg(Thy1-GCaMP6f)GP5.12Dkim/J |
025393 | C57BL/6J-Tg(Thy1-GCaMP6f)GP5.17Dkim/J |
028279 | C57BL/6J-Tg(Thy1-GCaMP6f)GP5.1Dkim/J |
028280 | C57BL/6J-Tg(Thy1-GCaMP6f)GP5.3Dkim/J |
024276 | C57BL/6J-Tg(Thy1-GCaMP6f)GP5.5Dkim/J |
7 strains use this protocol |