Quantitative real-time PCR assay to detect transgene copy number in cotton (Gossypium hirsutum).
A TaqMan quantitative real-time PCR detection system was developed to examine transgene copy number in cotton. GhUBC1, a gene validated to be present as a single copy per haploid Gossypium hirsutum genome, was used as the endogenous reference to estimate copy number of GFP and selection marker NPTII in 28 T0 plants. This system was found to be more accurate than genomic Southern blot hybridization and could effectively tell homozygotes from heterozygotes in a T1 transgenic cotton population. Therefore it is suitable for efficient and cost effective early screening of transgenic seedlings and identifying transgene homozygotes in segregation populations.
Yi
Cheng Xin
CX
Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604, Singapore.
Zhang
Jun
J
Chan
Ka Man
KM
Liu
Xiao Kun
XK
Hong
Yan
Y
D020090:Chromosome Segregation; D032461:Chromosomes, Plant / Q000235:genetics; D018628:Gene Dosage; D003368:Gossypium / Q000235:genetics*; D030821:Plants, Genetically Modified; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D019076:Transgenes / Q000235:genetics*