Analysis by quantitative PCR of zygosity in genetically modified organisms.
It is extremely useful to define a rapid and accurate method for identifying homozygous and heterozygous transgenic animals prior to setting up breeding programs for transgenic colonies and in experiments in which gene dosage effects could have a functional impact. Southern-blotting is a means of identifying zygosity, but such a method is time consuming and produces a high level of ambiguous results. Some years ago, we described the rapid, precise, non-ambiguous, and high-throughput identification of zygosity in transgenic animals by real-time PCR. This technique allows us to make a clear-cut identification of transgenic rats, transgenic mice, and double-transgenic pigs. Since 2002, however, several authors have made improvements to this method. The following paper describes the ease with which zygosity is determined using real-time PCR.
Tesson
Laurent
L
INSERM, UMR643, Nantes, France.
Rémy
Séverine
S
Ménoret
Séverine
S
Usal
Claire
C
Anegon
Ignacio
I
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D006580:Genetic Carrier Screening / Q000379:methods*; D006579:Heterozygote; D006720:Homozygote; D051379:Mice; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D051381:Rats