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This assay cannot distinguish hemi from hom
Tg= 63 bp
IPC = 74 bp
Tg Sequence: ACAAGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGTCTGGTGACCGTGACCCAGGACTCCTCCCTGcaggacggcacgctgatcTACAAGGTGAAGATGCGCGGCACCAACTTCCCCCCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGG
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
52208 | GTG ACC GTG ACC CAG GAC | Transgene Forward | A | |||
52209 | CGC GCA TCT TCA CCT TGT | Transgene Reverse | A | |||
52210 | Fluorophore-1 | CAG GAC GGC ACG CTG ATC | Quencher-1 | Tg Probe | tdTomato | |
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 | |
52208 | 0.40 uM |
52209 | 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 |
---|---|
035560 | B6(129S4)-Tg(tetO-Cxcl2,-tdTomato)#Forsb/Mmjax |
030219 | B6.Cg-Igs7tm139.1(tetO-EGFP,CAG-tdTomato,-tTA2)Hze/J |
037382 | B6.Cg-Igs7tm224(CAG-EGFP,CAG-dTomato)Tasic/J |
037383 | B6.Cg-Igs7tm227(CAG-EGFP,CAG-tdTomato)Tasic/J |
033125 | B6.FVB-Tg(tetO-Bcl2,-tdTomato)LM1Ryu/J |
036087 | B6;129-Gt(ROSA)26Sortm3Cos/J |
018936 | B6;C-Tg(Erv4-tdTomato)449Slp/J |
036272 | B6;CBA-Gt(ROSA)26Sorem1(CAG-Ikzf2,-tdTomato)Zyliu/J |
027395 | C57BL/6-Tg(Pvalb-tdTomato)15Gfng/J |
038819 | STOCK Gt(ROSA)26Sorem3(CAG-Insm1,-tdTomato)Zyliu/J |
038629 | STOCK Gt(ROSA)26Sortm65.2(CAG-tdTomato)Hze Pdgfrbtm1.1(flpo)Leol/LeolJ |
030292 | STOCK Ntrk3tm2.1Ddg/J |
038627 | STOCK Pdgfratm1(flpo)Leol Gt(ROSA)26Sortm65.2(CAG-tdTomato)Hze/LeolJ |
038628 | STOCK Pdgfratm2.1(flpo)Leol Gt(ROSA)26Sortm65.2(CAG-tdTomato)Hze/LeolJ |
026690 | STOCK Tg(Ucp1-luc2,-tdTomato)1Kajim/J |
15 strains use this protocol |