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This assay can be used to detect FLPe.
This assay cannot be used to detect FLPo.
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
17772 | TGC CGG TCC TAT TTA CTC GT | Transgene Forward | A | |||
17773 | TAC TTC TTT AGC GCA AGG GGT AG | Transgene Reverse | A | |||
oIMR8744 | CAA ATG TTG CTT GTC TGG TG | Internal Positive Control Forward | A | |||
oIMR8745 | GTC AGT CGA GTG CAC AGT TT | Internal Positive Control Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
17772 | 0.50 uM |
17773 | 0.50 uM |
oIMR8744 | 0.50 uM |
oIMR8745 | 0.50 uM |
Glycerol | 6.50 % |
Dye | 1.00 X |
Kapa 2G HS taq polym | 0.03 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 65.0 | -- | -0.5 C per cycle decrease |
4 | 68.0 | -- | |
5 | -- | repeat steps 2-4 for 10 cycles (Touchdown) | |
6 | 94.0 | -- | |
7 | 60.0 | -- | |
8 | 72.0 | -- | |
9 | -- | repeat steps 6-8 for 28 cycles | |
10 | 72.0 | -- | |
11 | 10.0 | -- | hold |
Stock Number | Strain Name |
---|---|
021072 | B6(Cg)-Slc6a8tm1.2Clar/J |
021828 | B6(SJL)-Lrrk2tm3.1Mjff/J |
025930 | B6(SJL)-Pik3cbtm1.1Jdo/J |
018445 | B6.129-Mre11atm1.1Rchd/J |
022364 | B6.129P2(SJL)-Ntrk3tm1Ddg/J |
018540 | B6.129S6(Cg)-Bpnt1tm2.1Yrk/J |
034712 | B6.Cg-Cyp27a1tm1.1Dpm/J |
021191 | B6.Cg-Pvalbtm2.1(flpe)Hze/J |
013717 | B6.Cg-Slc20a1tm1.2Cmg/J |
012627 | B6.Cg-Stard4tm1.1Bres/J |
012631 | B6.Cg-Tyrc-2J Sharpintm1.1Sun/J |
005703 | B6.Cg-Tg(ACTFLPe)9205Dym/J |
036551 | B6.FVB(Cg)-Tg(Pmch-cre)1Lowl/J |
012653 | B6;129-Pax7tm1.1Fan/J |
014168 | B6;129-Pot1atm1.1Tdl/J |
012623 | B6;129S4-Carftm1.1Aew/J |
016168 | B6;129S4-Fbxl3Gt(FHCRC-GT-S13-12G1)Sor/JtJ |
018353 | B6;129S4-Ror1tm1.1Meg/J |
034731 | B6N(129S4)-Srd5a3tm1c(EUCOMM)Wtsi/J |
022365 | C57BL/6-Il15ratm2.1Ama/J |
035185 | STOCK Adgrl1tm2c(EUCOMM)Hmgu/SudJ |
023199 | STOCK Brsk2tm2.1Jrs/J |
010702 | STOCK Gad2tm1(cre/ERT2)Zjh/J |
022972 | STOCK Pros1tm1Grl/J |
014099 | STOCK Tg(Pmch-cre)1Lowl/J |
25 strains use this protocol |