Determination of transgene copy number by real-time quantitative PCR.
Efficient methods to characterize transgenic plants are important to quickly understand the state of the transformant. Determining transgene copy number is an important step in transformant characterization and can differentiate between complex and simple transformation events. This knowledge can be extremely useful when determining what future experiments and uses the transgenic lines can be utilized for. The method described here uses real-time quantitative PCR to determine the transgene copy number present in the genome of the transformant. Specifically, this method measures the relative transgene copy number by comparing it with an endogenous gene with a known copy number. This method is a quick alternative to the Southern blot, a method that is commonly used to determine gene copy number, and is effective when screening large numbers of transformants.
Shepherd
Colin T
CT
AimsBio, Inc., USA. coshep@hotmail.com
Moran Lauter
Adrienne N
AN
Scott
M Paul
MP
D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics / Q000302:isolation & purification; D018628:Gene Dosage; D005821:Genetic Techniques; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods*; D003313:Zea mays / Q000235:genetics*
D017931:DNA Primers; D004274:DNA, Recombinant