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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate
instrument specific Fluorophore/Quencher combination. The transgene genotype is
determined by comparing ΔCt values of each unknown sample against known
homozygous and hemizygous controls, using appropriate endogenous references.
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
11287 | GGA GAC ACG GAA AGT GAA GG | Transgene Forward | A | |||
11288 | GGC CTC GCT CTG GTT GTA G | Transgene Reverse | A | |||
11289 | Fluorophore-1 | CCG AGT GGA CCT GGG GAC CC | 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 |
---|---|
ddH2O | |
Kapa Probe Fast QPCR | 1.00 X |
11287 | 0.40 uM |
11288 | 0.40 uM |
oIMR1544 | 0.40 uM |
oIMR3580 | 0.40 uM |
Tg Probe | 0.15 uM |
IC Probe | 0.15 uM |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 95.0 | -- | |
2 | 95.0 | -- | |
3 | 60.0 | -- | repeat steps 2-3 for 40 cycles |
Stock Number | Strain Name |
---|---|
004191 | B6.Cg-Immp2lTg(HLA-A/H2-D)2Enge/J |
003475 | C57BL/6-Mcph1Tg(HLA-A2.1)1Enge/J |
005512 | NOD.B6-Mcph1Tg(HLA-A2.1)1Enge/DvsJ |
006611 | NOD.Cg-B2mtm1Unc Tg(HLA-A/H2-D/B2M)1Dvs/DvsJ |
035844 | NOD.Cg-Flt3em1Akp Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(HLA-A/H2-D/B2M)1Dvs/J |
009617 | NOD.Cg-Mcph1Tg(HLA-A2.1)1Enge Prkdcscid Il2rgtm1Wjl/SzJ |
006609 | NOD.Cg-Mcph1Tg(HLA-A2.1)1Enge Prkdcscid/DvsJ |
006604 | NOD/ShiLtDvs-Tg(HLA-A/H2-D/B2M)1Dvs/J |
8 strains use this protocol |