Stock No: 005703
Protocol 22103: Probe Assay - Generic FLP
Version 6.0

Notes

The genotyping protocol(s) presented here have been optimized for reagents and conditions used by The Jackson Laboratory (JAX). To genotype animals, JAX recommends researchers validate the assay independently upon receipt of animals into their facility. Reaction cycling temperature and times may require additional optimization based on the specific genotyping reagents used.

Expected Results

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
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
oIMR8940 AGT AGT GAT CAG GTA TTG CTG TTA TCT G Forward A
oIMR8941 GAC TAA TGT TGT GGG AAA TTG GAG Reverse A
TmoIMR0105 Fluorophore-1 CCA ATG GTC GGG CAC TGC TCA A Quencher-1 IC Probe A
TmoIMR0126 Fluorophore-2 TAT ACG TTG TCC TGG CCA CGG CA Quencher-2 Tg Probe A

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
oIMR1544 0.40 uM
oIMR3580 0.40 uM
oIMR8940 0.40 uM
oIMR8941 0.40 uM
Wt Probe 0.15 uM
Mutant Probe 0.15 uM
DNA

Cycling

Step Temp °C Time Note
1 95.0 --
2 95.0 --
3 60.0 --
4 -- repeat steps 2-3 for 40 cycles
5 4.0 -- Forever
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

Strains Using This Protocol

Stock Number Strain Name
029290 B6(Cg)-Sgtatm1.1Drmcr/J
021072 B6(Cg)-Slc6a8tm1.2Clar/J
024469 B6(SJL)-Lrrk2tm3.1Mjfa/J
021828 B6(SJL)-Lrrk2tm3.1Mjff/J
018445 B6.129-Mre11atm1.1Rchd/J
022364 B6.129P2(SJL)-Ntrk3tm1Ddg/J
013085 B6.129S-Pbx1tm3.1Mlc/J
018540 B6.129S6(Cg)-Bpnt1tm2.1Yrk/J
012839 B6.129X1(Cg)-Tnfrsf4tm2(cre)Nik/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
008538 B6.Cg-Tg(Cspg4-cre/Esr1*)BAkik/J
012653 B6;129-Pax7tm1.1Fan/J
014168 B6;129-Pot1atm1.1Tdl/J
021443 B6;129S-Tgif1tm1Caw/J
012623 B6;129S4-Carftm1.1Aew/J
016168 B6;129S4-Fbxl3Gt(FHCRC-GT-S13-12G1)Sor/JtJ
018353 B6;129S4-Ror1tm1.1Meg/J
022407 B6;D2-Tg(Phox2b-flpo)3276Grds/J
026526 B6;SJL-Akap6tm1.1Mskf/Mmjax
034712 B6J.Cg-Cyp27a1tm1.1Dpm/J
034731 B6N(129S4)-Srd5a3tm1c(EUCOMM)Wtsi/J
022365 C57BL/6-Il15ratm2.1Ama/J
035185 STOCK Adgrl1tm2c(EUCOMM)Hmgu/SudJ
025665 STOCK Asxl1tm1.1Iaai/J
023199 STOCK Brsk2tm2.1Jrs/J
026199 STOCK F8tm1.1Rmnt/J
010702 STOCK Gad2tm1(cre/ERT2)Zjh/J
022972 STOCK Pros1tm1Grl/J
32 strains use this protocol