Protocol 37869: Probe Assay - Grm7<Tg(SMN2)89Ahmb>-Probe

FVB.Cg-Grm7^{Tg(SMN2)89Ahmb} Smn1^tm1Msd Tg(SMN2*delta7)4299Ahmb/J

Stock No: 005025

Version 1.0

Notes

Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.

 

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

>chr6:111099925+111100076 152bp CAAACCAAAGCTCCAGAATC GTGGCCAAATGTCAGTCACC

Mut= 158 bp

Wt= 152 bp

Fam=Mut

Hex=Wt

Sequence

Wt Sequence:

tctacattactgacctaccagggatgaggcattaggggaaatttttccaactcagtgtcatagcttctgtgtgctgtatgaactgataagttaacttttaacactgtataaattctgaaaacaaaccaaagctccagaatctttgaaaaatgagtttttgtttaagaaaccagaaatatgtagcctcccaaattgtcTGtaccctctgagttctcaattatttttaagtgtctgtgctacttagaggtggccaggtgactgacatttggccactatgaaggaagaaac

Mutant Sequence:

GTATAAATTCTGAAAACAAACCAAAGCTCCAGAATCTTTGAAAAATGAGTTTTTGTTTAAGAAACCAGAAATATGTAGCCTCCCAAATTgtctCAAAGTAATCTCTGTCTGATCCCTTAATGAAGGATCTCTTCAAACACAGCAACTTTTGTCTGTCTTCTGAACGTAGGTACAGCTGTGGCTG

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
52364		CAA ACC AAA GCT CCA GAA TC		Common	A
52365		GTG GCC AAA TGT CAG TCA CC		Wild type Reverse	A
52366		TGT ACC TAC GTT CAG AAG ACA GAC		Mutant Reverse	A
52367	Fluorophore-1	CCC AAA TTG TCT GTA CCC TCT GAG	Quencher-1	WT Probe
52368	Fluorophore-2	CCC TTA ATG AAG GAT CTC TTC AAA CAC	Quencher-2	MUT Probe

Reaction A

Component	Final Concentration
Kapa Probe Fast QPCR	1.00 X
ddH2O
52364	0.40 uM
52365	0.40 uM
52366	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
007222	B6.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN1*A2G)2023Ahmb/J
006964	B6.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN2*delta7)4299Ahmb/J
006773	B6.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd/J
006385	B6;129P2-Syt1^tm3Sud/J
006386	B6;129P2-Syt1^tm5Sud/J
008209	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(ACTA1-SMN)69Ahmb/J
007968	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN1*A2G)2023Ahmb/2J
005026	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN1*A2G)2023Ahmb/J
007952	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN2*delta7)4299Ahmb/2J
005025	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN2*delta7)4299Ahmb/J
031906	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN2-SMN1*T274I)5Ahmb/J
007949	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd/2J
005024	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd/J
030214	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(Prnp-PLS3)14Ahmb Tg(SMN2*delta7)4299Ahmb/J
030970	FVB.Cg-Mapk10^tm1Flv Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN2*delta7)4299Ahmb/LganJ
031907	STOCK Grm7^Tg(SMN2)89Ahmb Tg(SMN2-SMN1*Q282A)2Ahmb Smn1^tm1Msd/J
008203	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(ACTA1-SMN)63Ahmb/J
032003	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(Hlxb9-GFP)1Tmj Tg(SMN2*delta7)4299Ahmb/J
006570	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(Hlxb9-GFP)1Tmj/J
008212	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(Prnp-SMN)92Ahmb/J
025102	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(Rnu7-SMN2*,PGK1-EGFP)4Dasch/DaschMmjax
018916	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN1-SMN2*)16Cll/CllJ
031908	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN2-SMN1*D44V)10Ahmb/J
008783	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm3(SMN2/Smn1)Mrph Tg(CAG-cre/Esr1*)5Amc Tg(SMN2*delta7)4299Ahmb/J
007951	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm3(SMN2/Smn1)Mrph Tg(SMN2*delta7)4299Ahmb/J
007022	STOCK Mnx1^tm4(cre)Tmj Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN2*delta7)4299Ahmb/J
017600	STOCK Tg(tetO-SMN2,-luc)#bAhmb/J
27 strains use this protocol