Protocol 6688 - Tg(WapMyc)212Bri

Notes

This assay will NOT distinguish hemizygous from homozygous transgenic animals.

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

Transgene = 58 bp

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
12485		GCC CAA GTT GGG AAG ACA AC		Transgene Forward
12486		CCA GCT TGG TTC CCG AAT AGA C		Transgene Reverse

Reaction A

Component	Final Concentration
ddH2O
AB PCR BufferII	1.00
MgCl2	2.00
dNTP	0.20
	0.00
	0.00
	0.00
	0.00
DNA Loading Dye	0.69
Taq DNA Polymerase	0.03
DNA

Cycling

Step	Temp °C	Time	Note
1	94.0	--
2	94.0	--
3		--
4	72.0	--
5		--	Go to step 2-4, 35 times
6	72.0	--
7	10.0	--	hold

JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

Strains Using This Protocol

This is the only strain that uses this protocol.