Protocol 22103: Probe Assay - Generic FLP

B6;SJL-Akap6^tm1.1Mskf/Mmjax

Stock No: 026526

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	40.0	--	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)-Sgta^tm1.1Drmcr/J
021072	B6(Cg)-Slc6a8^tm1.2Clar/J
021828	B6(SJL)-Lrrk2^tm3.1Mjff/J
018445	B6.129-Mre11a^tm1.1Rchd/J
022364	B6.129P2(SJL)-Ntrk3^tm1Ddg/J
013085	B6.129S-Pbx1^tm3.1Mlc/J
018540	B6.129S6(Cg)-Bpnt1^tm2.1Yrk/J
034712	B6.Cg-Cyp27a1^tm1.1Dpm/J
024469	B6.Cg-Lrrk2^tm3.1Mjfa/J
021191	B6.Cg-Pvalb^tm2.1(flpe)Hze/J
013717	B6.Cg-Slc20a1^tm1.2Cmg/J
012627	B6.Cg-Stard4^tm1.1Bres/J
012631	B6.Cg-Tyr^c-2J Sharpin^tm1.1Sun/J
005703	B6.Cg-Tg(ACTFLPe)9205Dym/J
036551	B6.FVB(Cg)-Tg(Pmch-cre)1Lowl/J
012653	B6;129-Pax7^tm1.1Fan/J
014168	B6;129-Pot1a^tm1.1Tdl/J
021443	B6;129S-Tgif1^tm1Caw/J
012623	B6;129S4-Carf^tm1.1Aew/J
016168	B6;129S4-Fbxl3^Gt(FHCRC-GT-S13-12G1)Sor/JtJ
018353	B6;129S4-Ror1^tm1.1Meg/J
022407	B6;D2-Tg(Phox2b-flpo)3276Grds/J
026526	B6;SJL-Akap6^tm1.1Mskf/Mmjax
022365	C57BL/6-Il15ra^tm2.1Ama/J
040836	C57BL/6J-Cux1^em1(flpo)Zjh/J
040838	C57BL/6J-Igfn1^em1(flpo)Zjh/J
040837	C57BL/6J-Wfs1^em1(flpo)Zjh/J
035185	STOCK Adgrl1^tm2c(EUCOMM)Hmgu/SudJ
025665	STOCK Asxl1^tm1.1Iaai/J
023199	STOCK Brsk2^tm2.1Jrs/J
026199	STOCK F8^tm1.1Rmnt/J
010702	STOCK Gad2^tm1(cre/ERT2)Zjh/J
022972	STOCK Pros1^tm1Grl/J
014099	STOCK Tg(Pmch-cre)1Lowl/J
34 strains use this protocol