Protocol 22178: QPCR Assay - iCre QPCR

B6.Cg-Hprt1^{tm354(Ple283-icre/ERT2)Ems}/Mmjax

Stock No: 023699

Version 4.0

Notes

Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination. The transgene genotype is determined by comparing ΔCt values of each unknown sample against known homozygous and hemizygous controls, using appropriate endogenous references.

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
17817		ACA ACT ACC TGT TCT GCC G		Forward	A
17818		GCC TCA AAG ATC CCT TCC AG		Reverse	A
17819	Fluorophore-1	TTC TGC CAC CTC CCA ACT GTC C	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

Reaction A

Component	Final Concentration
ddH2O
Kapa Probe Fast QPCR	1.00 X
17817	0.40 uM
17818	0.40 uM
oIMR1544	0.40 uM
oIMR3580	0.40 uM
Tg Probe	0.15 uM
IC Probe	0.15 uM
DNA

Cycling

Step	Temp °C	Time	Note
1	95.0	--
2	95.0	--
3	60.0	--	repeat steps 2-3 for 40 cycles

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
035764	B6(129)-Jchain^em1(icre/ERT2)Deep/J
039826	B6(129S4)-Fcgr1^em1.1(icre)Moli/J
035435	B6(129S4)-Xcr1^tm1.1(cre)Kmm/J
009587	B6(129S4)-Et(icre)1402Rdav/J
009588	B6(129S4)-Et(icre)1470Rdav/J
009589	B6(129S4)-Et(icre)1555Rdav/J
009586	B6(129S4)-Et(icre)754Rdav/J
010696	B6(129S4)-Et(icre/ERT2)10596Rdav/J
010697	B6(129S4)-Et(icre/ERT2)10727Rdav/J
012689	B6(129S4)-Et(icre/ERT2)14163Rdav/J
012690	B6(129S4)-Et(icre/ERT2)14208Rdav/J
012694	B6(129S4)-Et(icre/ERT2)14915Rdav/J
012687	B6(129S4)-Tg(SYN1-icre/mRFP1)9934Rdav/J
038536	B6(Cg)-Tg(Cd8a-icre/ERT2,-GFP)#Daav/J
038538	B6(Cg)-Tg(Zbtb7b/CD4-icre/ERT2,-CD2)13Daav/J
039650	B6(SJL)-Cd40lg^tm1.1(icre)Tega/J
038129	B6.Cg-Ascl3^tm2.1(icre/ESR1*)Ovi/J
038342	B6.Cg-Gas1^em1(icre/ERT2)Nono/J
023677	B6.Cg-Hprt1^tm332(Ple272-icre/ERT2)Ems/Mmjax
023690	B6.Cg-Hprt1^tm345a(icre/ERT2)Ems/Mmjax
023691	B6.Cg-Hprt1^tm346a(Ple264-icre/ERT2)Ems/Mmjax
023692	B6.Cg-Hprt1^tm347a(Ple155-icre/ERT2)Ems/Mmjax
023693	B6.Cg-Hprt1^tm348a(Ple265-icre/ERT2)Ems/Mmjax
023694	B6.Cg-Hprt1^tm349a(hs671-icre/ERT2)Ems/Mmjax
023695	B6.Cg-Hprt1^tm350a(Ple232-icre/ERT2)Ems/Mmjax
023696	B6.Cg-Hprt1^tm351a(Ple198-icre/ERT2)Ems/Mmjax
023697	B6.Cg-Hprt1^tm352a(Hspa1a-icre/ERT2)Ems/Mmjax
023698	B6.Cg-Hprt1^tm353(Ple274-icre/ERT2)Ems/Mmjax
023699	B6.Cg-Hprt1^tm354(Ple283-icre/ERT2)Ems/Mmjax
023705	B6.Cg-Hprt1^tm360a(Ple253-icre/ERT2)Ems/Mmjax
023706	B6.Cg-Hprt1^tm361a(Ple267-icre/ERT2)Ems/Mmjax
023708	B6.Cg-Hprt1^tm363a(hs1218-icre/ERT2)Ems/Mmjax
023710	B6.Cg-Hprt1^tm365a(Ple266-icre/ERT2)Ems/Mmjax
023711	B6.Cg-Hprt1^tm366a(Ple67-icre/ERT2)Ems/Mmjax
023712	B6.Cg-Hprt1^tm367a(Ple251-icre/ERT2)Ems/Mmjax
039341	B6.Cg-Klf1^em1(icre)Jbkr/J
038823	B6.Cg-Myh11^em1(icre)Jifz/J
015850	B6.Cg-Pde6b^+ Tg(Rho-icre)1Ck/Boc
040344	B6.Cg-Slc6a9^em1(icre)Mira/J
026516	B6.Cg-Tg(Avil-icre/ERT2)AJwo/J
008520	B6.Cg-Tg(CD2-icre)4Kio/J
036935	B6.FVB(Cg)-Tp(X)1Soff^Tg(Myh11-icre/ERT2)1Soff/ZjngJ
019079	B6.FVB-Tg(Myh11-icre/ERT2)1Soff/J
017490	B6.FVB-Tg(Stra8-icre)1Reb/LguJ
030663	B6;129S-Gcg^tm1.1(icre)Gkg/J
040784	B6;129S6-Pgr^em1(icre/ERT2)Fjd/J
038989	B6;C3-Oxtr^em4(icre/ERT2)Yinn/2J
025809	B6;CBA-Tg(Fgfr3-icre/ERT2)4-2Wdr/J
024768	B6;CBA-Tg(Upk2-icre/ERT2)1Ccc/J
031645	B6;D2-Tn(pb-CAG-dcas9,-EGFP)1Yangh/J
028547	B6;SJL-Tg(Cyp17a1-icre)AJako/J
040426	B6J(B6N)-Tg(Nr4a1-EGFP/cre)820Khog/PalcaJ
036235	B6N.Cg-Tg(Cxcl12-icre/ERT2)L289Nono/J
035229	C57BL/6-Ccr2^em1(icre/ERT2)Peng/J
025124	C57BL/6-Tg(Adipoq-icre/ERT2)1Soff/J
040204	C57BL/6-Tg(Amelx-icre)872Ppap/JdbJ
038650	C57BL/6-Tg(Crhr1-icre,-tdTomato)3Njus/J
038197	C57BL/6-Tg(Nmur1-iCre,-eGFP)1Dart/J
016617	C57BL/6-Tg(Nr4a1-EGFP/cre)820Khog/J
016583	C57BL/6-Tg(Slc6a3-icre/ERT2)2Gloss/J
040787	C57BL/6J-Cd3g^em1(icre)Xhsu/J
040610	C57BL/6J-Kremen1^em1(icre)Cai/J
041041	C57BL/6J-Mog^em1(icre)Lusu/J
016582	C57BL/6N-Tg(Slc32a1-icre/ERT2)3Gloss/J
030676	C57BL/6N-Tg(Tg-cre/ERT2)1Kero/J
019098	FVB-Tg(Csf1r-cre/Esr1*)1Jwp/J
021024	FVB-Tg(Csf1r-icre)1Jwp/J
039016	NOD.Cg-H2^b Commd10^Tg(Vav1-icre)A2Kio/JkraJ
008694	NOD/ShiLt-Tg(Foxp3-EGFP/cre)1cJbs/J
037019	STOCK Foxp3^tm4(icre)Tch/Mmjax
034261	STOCK Ighg3^tm1.1(icre)Gbrt/J
037139	STOCK Myl1^tm1(cre)Sjb Atg7^tm1Tchi Gaa^tm1Rabn/PuroMmjax
039298	STOCK Shox2^em1(icre)Zyliu/J
012692	STOCK Et(icre/ERT2)14602Rdav/J
012693	STOCK Et(icre/ERT2)14624Rdav/J
008208	STOCK Tg(Stra8-icre)1Reb/J
030061	STOCK Tg(Tbx18-icre)3Fech/J
016584	STOCK Tg(Tph2-icre/ERT2)6Gloss/J
78 strains use this protocol