Protocol 21935: End Point Analysis Assay - Crb1<rd8>End Point

B6.Cg-Crb1^rd8 Jak3^m1J/BocJ

Stock No: 005558

Version 3.0

Notes

Mutant = -/-  B6N (mutation fixed on this background)
Heterozygote = C/-
Wild type = C/C B6J

Use controls from DNA Resorces jr 3392

Mutation is a deleted C

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

Mutant = -/-
Heterozygote = C/-
Wild type = C/C

Mutation is a deleted C

 

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
16217		TTG CAT GGA GGA AAC TGT GA		Forward	A
16218		TGT ATC CAG GCT CAC ACC TG		Reverse	A
16219	Fluorophore-1	CAG TTC TTA TCG GTG TGC CTG	Quencher-1	WT Probe
16220	Fluorophore-2	CAG TTC TTA TGG TGT GCC TG	Quencher-2	MUT Probe

Reaction A

Component	Final Concentration
Kapa Probe Fast QPCR	1.00 X
ddH2O
16217	0.40 uM
16218	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
030931	129S6.Cg-Pex1^tm1.1Sjms/Mmjax
034736	B6(Cg)-Slc17a8^em1(icre/ERT2)Zyliu/J
019141	B6.129S6(FVB)-S1pr1^tm2.1Rlp/J
039326	B6.Cg-Crb1^rd8 Arhgef12^Tvrm266/PjnMmjax
005558	B6.Cg-Crb1^rd8 Jak3^m1J/BocJ
031293	B6.Cg-Crb1^rd8 Prkcq^tm1Litt/JBoc
008451	B6.Cg-Ptprc^a Cx3cr1^tm1Litt/LittJ
004852	B6;129-Crb1^rd8/J
031229	C57BL/6J-Ctnna1^Tvrm5/PjnMmjax
005304	C57BL/6NJ
000654	CBA/CaJ
003392	STOCK Crb1^rd8/J
013169	STOCK Nphp1^tm1Jgg/J
13 strains use this protocol