Protocol 16761: Sanger sequencing Assay - Foxn1<em1Dvs>

NOD.Cg-Foxn1^em1Dvs Prkdc^scid Il2rg^tm1Wjl/J

Stock No: 026263

Version 1.0

Notes

Mut = X

WT= Y

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

Sequence

SEQ

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
24362		AGT GTG GAT CTC TAT CCA AGT CC		Forward
24363		CAG CAG CAT CTA CTG AGT CCA		Reverse

Reaction A

Component	Final Concentration
ddH2O
Kapa 2G HS buffer	1.00
MgCl2	2.00
dNTPS-kapa	0.20
Primer 1	0.50
Primer 2	0.50
Glycerol	5.00
Kapa 2G HS taq polym	0.01
DNA

Cycling

Step	Temp °C	Time	Note
1	94.0	--
2	94.0	--
3	65.0	--	-0.5 C per cycle decrease
4	68.0	--
5		--	repeat steps 2-4 for 10 cycles
6	94.0	--
7	60.0	--
8	72.0	--
9		--	repeat steps 6-8 for 28 cycles
10	72.0	--
11	10.0	--	hold

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
026263	NOD.Cg-Foxn1^em1Dvs Prkdc^scid Il2rg^tm1Wjl/J
029390	NOD.Cg-Foxn1^em1Dvs/J
2 strains use this protocol