Protocol 31920: Probe Assay - Generic LacZ Probe
Version 5.0

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
37301 GTT TAT GCA GCA ACG AGA CG Transgene Reverse A
37302 Fluorophore-1 CAG GAT ATG TGG CGG ATG AGC Quencher-1 Tg Probe
mom0095 TGA CGG CAG TTA TCT GGA AG Transgene Forward A
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
Kapa Probe Fast QPCR 1.00 X
ddH2O
37301 0.40 uM
mom0095 0.40 uM
oIMR1544 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
012668 129S.129-Lrp5tm1Grw/J
038651 129S1.Cg-Tg(Hmgcr-lacZ)H253Sest/EmsJ
002957 B6.129-Dll1tm1Gos/J
004276 B6.129-Figntm1Frk/Frk
004478 B6.129-Foxd1tm1Lai/J
003870 B6.129-Plin1tm1Chan/J
019146 B6.129-Tmc1tm1.1Ajg/J
030985 B6.129P2(Cg)-Aph1atm1Bdes Aph1btm1Bdes Aph1ctm1Bdes/J
009348 B6.129P2(Cg)-Hprt1tm17(Ple48-lacZ)Ems/Mmjax
012575 B6.129P2(Cg)-Hprt1tm39(Ple24-lacZ)Ems/Mmjax
012333 B6.129P2(Cg)-Hprt1tm45(Ple67-lacZ)Ems/Mmjax
012733 B6.129P2(Cg)-Hprt1tm53(CAG-lacZ)Ems/Mmjax
012347 B6.129P2(Cg)-Hprt1tm64(Ple170-lacZ)Ems/Mmjax
012583 B6.129P2(Cg)-Hprt1tm68(Ple127-lacZ)Ems/Mmjax
012656 B6.129P2(Cg)-Hprt1tm70(Ple240-lacZ)Ems/Mmjax
012657 B6.129P2(Cg)-Hprt1tm71(Ple155-lacZ)Ems/Mmjax
012659 B6.129P2(Cg)-Hprt1tm73(Ple142-lacZ)Ems/Mmjax
012734 B6.129P2(Cg)-Hprt1tm74(Ple232-lacZ)Ems/Mmjax
018054 B6.129P2(Cg)-Pitx2tm1.1Dmm/J
013144 B6.129P2-Aim2Gt(CSG445)Byg/J
005801 B6.129P2-Esrratm1Dgen/J
005814 B6.129P2-Grm1tm1Dgen/J
034597 B6.129P2-Kif13aGt(AF0553)Wtsi/AhcJ
030421 B6.129P2-Mbd2tm1Bh/JoezJ
023209 B6.129S-Sfrp1tm1Jrbn/PcmaJ
004950 B6.129S1-Syt7tm1Nan/J
036325 B6.129S6-Mecomtm1Aspe/Mmjax
005876 B6.CAST-(rs4222996-rs13476494)/LusJ
019547 B6.Cg-Cacna1ctm2Itl/J
017762 B6.Cg-Cacna1ftm1.2Sdie/J
030065 B6.Cg-Serpinf1tm1Craw/J
033441 B6.Cg-Slc15a1tm1Dgen Tg(SLC15A1)1Dsmi/Mmjax
006477 B6.Cg-Tg(CAG-lacZ-WGA)330Bbm/J
033345 B6.Cg-Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik/J
032889 B6.Cg-Tg(Cdh5-RHO/Adrb2,-lacZ)B27-3Mik/J
006229 B6.Cg-Tg(DRE-lacZ)2Gswz/J
033344 B6.Cg-Tg(Hcn4-COP4*L132C,-lacZ)B17-2Mik/J
024050 B6.Cg-Tg(Hmgcr-lacZ)H253Sest/J
023757 B6.Cg-Tg(tetO-tetX,lacZ)1Gogo/UmriJ
028353 B6.Cg-Tg(Vil1-EGFP)106Gum/J
018625 B6.FVB-Tg(Fabp4-lacZ)4Mosh/J
025740 B6;129-Prg4tm2Mawa/J
008704 B6;129-Smn1tm6(SMN2)Mrph/J
006251 B6;129-Tor1atm1Wtd/J
025197 B6;129-Vmn1r51tm1Dlc/J
025196 B6;129-Vmn1r51tm2.1Dlc/J
017524 B6;129-Tg(CMV-Bgeo,-WGA,-ALPP)1Mgmj/J
038621 B6;129P2-Ctcftm2.2Gal/J
024288 B6;129P2-Faptm2Schn/J
006744 B6;129P2-Gt(ROSA)26Sortm1Mom/MomJ
032534 B6;129P2-Maltm1Nsw/J
006665 B6;129P2-Or8a1tm13(Olr226)Mom/MomJ
012850 B6;129P2-TardbpGt(RRB030)Byg/J
026012 B6;129P2-Uts2rtm1Djbe/J
006465 B6;CBA-Tg(CAG-lacZ-WGA)330Bbm/J
007972 B6;CBA-Tg(Olfr151-taulacZ)4Mom/MomJ
006680 B6;CBA-Tg(Olfr16*,taulacZ)19Mom/MomJ
006672 B6;CBA-Tg(Olfr16*,taulacZ)7Mom/MomJ
007973 B6;CBA-Tg(Olfr16-taulacZ)1Mom/MomJ
007974 B6;CBA-Tg(Olfr160-taulacZ)V4-7Mom/MomJ
007976 B6;CBA-Tg(Olfr713-taulacZ)4Mom/MomJ
007975 B6;CBA-Tg(OR8A1-taulacZ)1Mom/MomJ
004141 B6;CBA-Tg(UAS-lacZ)65Rth/J
008344 B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J
018627 B6;SJL-Tg(Myl1-lacZ)1Ibdml/J
002621 B6;SJL-Tg(tetop-lacZ)2Mam/J
003299 B6;SWJ-Tg(TIMP3-lacZ)7Jeb/J
037014 B6J.129P2-Pofut2Gt(RST434)Byg/BchMmjax
024866 B6J.B6N(Cg)-Sec24ctm1a(EUCOMM)Wtsi/J
034731 B6N(129S4)-Srd5a3tm1c(EUCOMM)Wtsi/J
030130 B6N.129-Dkk2tm1Dwu/J
030043 B6N;129-LrbaGt(OST69142)Lex/WgkJ
002754 C57BL/6-Tg(LacZpl)60Vij/J
023048 C57BL/6-Tg(Peg3-lacZ)26Sass/J
027898 C57BL/6J-Tg(CAG-lacZ,-FUS,-EGFP)629Gyu/J
037062 FVB.129S-Neu1tm1Adz/J
005025 FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2*delta7)4299Ahmb/J
031323 FVB/N-Tg(Bmp5,Hspa1b-lacZ)639Kng/J
031325 FVB/N-Tg(Bmp5-Hmgcll1,Hspa1b-lacZ)1Kng/J
031324 FVB/N-Tg(Bmp5-Hmgcll1,Hspa1b-lacZ)665Kng/J
026047 FVB/N-Tg(Isl1-lacZ)5Cka/Mmjax
002856 FVB/N-Tg(TIE2-lacZ)182Sato/J
035185 STOCK Adgrl1tm2c(EUCOMM)Hmgu/SudJ
026229 STOCK Akap12tm1Ihg Rb1tm2Brn Tg(Pbsn-cre)4Prb/J
035392 STOCK Gsctm2Bhr/J
024284 STOCK Hcn4tm3Sev/J
013764 STOCK Hprt1tm57(Ple26-lacZ)Ems/Mmjax
012354 STOCK Hprt1tm66(Ple5-lacZ)Ems/Mmjax
012584 STOCK Hprt1tm69(Ple134-lacZ)Ems/Mmjax
013087 STOCK Mir7-2tm1Mtm/Mmjax
006578 STOCK Myoz2tm1Eno/J
006686 STOCK Or6b9tm1Mom/MomJ
006613 STOCK Tg(CAG-Bgeo,-Tle1,-ALPP)1Lbe/J
004623 STOCK Tg(TCF/Lef1-lacZ)34Efu/J
005728 STOCK Tg(tetO-Ipf1,lacZ)958.1Macd/J
95 strains use this protocol