Methodology

Design of an improved set of oligonucleotide primers for genotyping MeCP2^tm1.1Bird KO mice by PCR

Julie Miralvès^* , Eddy Magdeleine^* and Etienne Joly

Equipe de Neuro-Immuno-Génétique Moléculaire, IPBS, UMR CNRS 5089, 205 route de Narbonne, 31077 Toulouse Cedex, France

author email corresponding author email* Contributed equally

Molecular Neurodegeneration 2007, 2:16doi:10.1186/1750-1326-2-16

Published:	31 August 2007

Abstract

Background

The strain of MeCP2^tm1.1Bird mice is a broadly used model for Rett syndrome. Because males carrying the invalidated MeCP2 locus are sterile, this strain has to be maintained in a heterozygous state. All animals therefore have to be genotyped at every generation to discriminate those carrying the invalidated allele (+/- females and y/- males) from those that do not. This is conveniently carried out by PCR on tail genomic DNA but because the primer pairs described initially for this purpose yield very similar size DNA bands on the WT and the KO alleles, this requires to carry out two independent PCR reactions on tail DNA preparations from all animals.

Results

After cloning and sequencing the PCR fragment amplified on the KO allele, we tested several sets of primers that were designed to yield PCR fragments of different sizes on the KO and WT alleles.

Conclusion

We have thus identified a set of three primers that allows for efficient genotyping of the animals by a single PCR reaction. Furthermore, using of this set of primers also resolves a recurrent problem related to the tendency of one of the initial primers to give rise to a non specific band because of its capacity to anneal at both ends of a repeated genomic element which we have identified as MurvyLTR.