TCAAACTATTATCTTAAGCAAGTAGCTGTGGATATAAACTATGTCTCATTAATTTTATTTTAATACTTACCACAGCAATACCTCAAGATACCAAAACACATGGCCAACTGTGGGAATATTCTGATAGAAGAGAATATTATTGGGTAGTCTTCAGATCAGTGGGATAACTAGCCAAAAAGAATCTATGAAAGAAATCTAATCTAAATGATCCAGGATAGGTCTAAGAACTTCACTGAGATTTTATTAATTCACAATTAATCAAAAGATAGGTATTCATGATTCAAGCTGGCTATGTAAATACTTCTTTTCAGTGTCTTATTTTTCCCATTATAACTTTTTCTGTTTTTAACAGTTCCAGAAACGCCTAAGCCTAGTTGTGGGGATCCAGCTGCAAGAAAAACGTTAGTCTCTTGGCCTGAGCCTG(T/C)ATCTAAACCTGA(G/T)(T/C)CTGCATCTAAACCCCATGGATATGTTTTATGCTATAAGAACAATTCAGGTAATGTAAAATTCCACTAGGGAAACAAAATCAAGAATTTTTAAATGTTGTAATTATTGTTTTAGCCAGGGAGCTACAAGGCTTTTATGAAGGTGATGAGAGATTCATGTTTGAGTTTGCTTAGATGAACACGCATCTCATGGAGAAGAATAAATATCTAGATCATTTATTAAAAATAGTGGACAGGACTAGAGTCCTTTAGAGCAGGTTACTCATCAGGACATTCCTTTATCTCTGGGCTAAGGTGGGAATGTACTTAGCTCTCATAGA
Twelve nucleotide differences between the a and b alleles have been identified. This assay is designed to detect a T to C point mutation (indicated in red), however, additional nucleotide differences in close proximity have been indicated in green.
Mutant (Ptprc<a>)(CD45.1, Ly5.1) = C/C
Heterozygote = T/C
Wild type (Ptprc<b>)(CD45.2, Ly5.2) = T/T
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
15560 | CGC CTA AGC CTA GTT GTG G | Forward | A | |||
15561 | ATT CTT GAT TTT GTT TCC CTA GTG G | Reverse | A | |||
25560 | Fluorophore-1 | CCT GAG CCT GTA TCT AAA CCT GA | Quencher-1 | WT Probe | ||
25561 | Fluorophore-2 | CCT GAG CCT GCA TCT AAA CCT G | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
15560 | 0.40 uM |
15561 | 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 | -- | repeat steps 2-3 for 40 cycles |
Stock Number | Strain Name |
---|---|
007594 | B6.129P2-Ptprca Ightm1Mnz/J |
028269 | B6.129S6(C)-Inpp5dtm1.1Wgk/J |
006183 | B6.Cg-Col4a5tm1Yseg/J |
006259 | B6.Cg-Pepcb Ptprca Tg(UBC-scFv)2Nemz/J |
008451 | B6.Cg-Ptprca Cx3cr1tm1Litt/LittJ |
013198 | B6.Cg-Ptprca Mir223tm1Fcam/J |
037061 | B6.Cg-Ptprca Pepcb Tg(EEF1A1-IL15RA)1Nsat/J |
030450 | B6.Cg-Ptprca Pepcb Tg(TcrLCMV)1Aox/PpmJ |
039112 | B6.Cg-Ptprca Pepcb KitW-41J Gpi1a/IslabMmjax |
027509 | B6.Cg-Ptprca Pepcb Tg(TcraBC10,TcrbBC10)3Chi/J |
022486 | B6.Cg-Ptprca Tg(UBC-PA-GFP)1Mnz/J |
002014 | B6.SJL-Ptprca Pepcb/BoyJ |
007203 | B6;129S4-Zfand5Gt(ROSA)72Sor/J |
025854 | B6;FVB-Ptprca Tg(CAG-luc,-GFP)L2G85Chco Thy1a/J |
017351 | BKa.Cg-Ptprcb Bmi1tm1Ilw Thy1a/J |
007686 | BKa.Cg-Sox17tm2Sjm Ptprcb Thy1a/J |
018869 | C.129S4(Cg)-Il13tm2.1Lky/J |
006584 | CByJ.SJL(B6)-Ptprca/J |
035855 | NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg(HLA-A/H2-D/B2M)Dvs Tg(HLA-DRA,HLA-DRB1*0401)39-2Kito/J |
025855 | STOCK Ptprca Lag3tm1Doi Tg(CAG-luc,-GFP)L2G85Chco Thy1a/J |
037371 | STOCK Ptprca Pepcb H2u H2-T18a Tg(CD2-TcraAc1-9,-TcrbAc1-9)4Kio/WdcJ |
030113 | STOCK Ptprca Pepcb Pkmtm1.1Mgvh/J |
028062 | STOCK Ptprca TcrbLn1Bdb TcraLn1Bdb H2d/J |
23 strains use this protocol |