Stock No: 005110
Protocol 734: Restriction Enzyme Digest Assay - Tg(Sod1*G86R)M1Jwg
Version 1.1

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

Transgene = ~90 bp and ~55 bp
Internal Positive Control = ~145 bp
Separated by gel electrophoresis on a 5.0% agarose gel.**

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
oIMR3505 GAC ATC ATT GTT CAT CC Common
oIMR3506 AAT GAT GGA ATG CAC TCC TGA Common

Reaction A

Component Final Concentration
ddH2O
AB PCR BufferII 1.00 X
MgCl2 2.00 mM
dNTP 0.20 mM
oIMR3505 1.00 uM
oIMR3506 1.00 uM
DNA Loading Dye 0.69 mM
Taq DNA Polymerase 0.03 U/ul
DNA

Cycling

Step Temp °C Time Note
1 94.0 --
2 94.0 --
3 60.0 --
4 72.0 -- repeat steps 2-4 for 40 cycles
5 72.0 --
6 10.0 -- hold**
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

Reaction B

Component Final Concentration
AB PCR BufferII X
ddH2O
FspI

Strains Using This Protocol

This is the only strain that uses this protocol.