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This assy cannot distinguish hemi from hom
Tg= 128 bp
IPC = 74 bp
Fam=Mut
Cy5=IPC
Assay cannot distinguish hemi from hom
Tg Sequence: GATTAGATAAAAGTAAAGTGATTAACaGCGCATTAGAGcTGCTTAATGaGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTtGGTGTAGAGCAGCCTACAcTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAgCcATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAaAAGGGGAAAGCTGGCAAGATTTTTTACGcAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCaATGGA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
40699 | GGT TTA ACA ACC CGT AAA CTC G | Transgene Forward | A | |||
40700 | TGA GTA TGG TGC CTA TCT AAC ATC | Transgene Reverse | A | |||
40701 | Fluorophore-1 | CAG CCT ACA CTG TAT TGG CAT G | 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 | |
40699 | 0.40 uM |
40700 | 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 |
---|---|
023917 | B6.Cg-Lin28btm1.1Gqda/J |
034533 | B6.FVB-Tg(EmuSR-tTa)83Bop/DwfJ |
012433 | B6;C3-Tg(ACTA1-rtTA,tetO-cre)102Monk/J |
024851 | B6N.129S6-Sod2tm1(rtTA*M2,tetO)Jsg/J |
034802 | FVB/N-Tg(MMTV-tTA)25754Kuw/Mmjax |
017393 | FVB;B6-CriptTn(sb-rtTA)2427.SB1Ove/Mmjax |
005734 | NOD/Lt-Tg(Ins2-rtTA)1Ach/AchJ |
006999 | STOCK Dbttm1Geh Tg(Cebpb-tTA)5Bjd Tg(tetO-DBT)A1Geh/J |
032910 | STOCK Tg(BEST1-rtTA,tetO-cre)1Yzl/J |
024854 | STOCK Tg(Camk2a-tTA)1Mmay Fgf14Tg(tetO-MAPT*P301L)4510Kha/J |
003272 | STOCK Tg(Cebpb-tTA)5Bjd/J |
003273 | STOCK Tg(CMV-rtTA)4Bjd/J |
025672 | STOCK Tg(tetO-Fgfr2b/Igh)1.3Jaw/J |
026194 | STOCK Tg(Wnt1-rtTA2S*M2)1Whsu/J |
14 strains use this protocol |