Real-time PCR method using capturing oligo-immobilized PCR tubes to determine the specific gene for soybean and genetically modified soybean in food matrices.
A new real-time PCR method using capturing oligo-immobilized PCR tubes is described. This method was used to detect specific genes for soybean and genetically modified (GM) soybean in food matrices. In a standard reaction using soybean genomic DNA and a capturing oligo for the lectin gene (Le1) immobilized on the tube, we examined the effects of such hybridization conditions as the location, length, and amount of the capturing oligo, and the incubation time and temperature. Under optimized conditions, the copy number of Le1 was determined in a concentration-dependent manner from soybean genomic DNA and soybean lysate (DNA 10-1000 ng, r=0.99; lysate 1-100%, r=0.99). The copy number of a Roundup Ready soybean (RRS) gene was also successfully detected in a concentration-dependent manner (1-100%, r=0.99) from GM soybean lysate, using PCR tubes with an immobilized capturing oligo for the transgene. Our data indicate that this is a rapid and simple method to determine specific genes for soybean and GM soybean in food matrices.
Harikai
Naoki
N
School of Pharmaceutical Sciences, Mukogawa Women's University, Hyogo, Japan.
Saito
Shin
S
Abe
Midori
M
Kondo
Kazunari
K
Kitta
Kazumi
K
Akiyama
Hiroshi
H
Teshima
Reiko
R
Kinoshita
Kenji
K
D001483:Base Sequence; D018744:DNA, Plant / Q000235:genetics; D030841:Food, Genetically Modified; D018628:Gene Dosage; D017343:Genes, Plant / Q000235:genetics*; D005998:Glycine / Q000031:analogs & derivatives; D006358:Hot Temperature; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D037121:Plant Lectins / Q000235:genetics; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods*; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000235:genetics*; D013045:Species Specificity; D013696:Temperature; D013997:Time Factors; D019076:Transgenes; D000097797:Glyphosate
D018744:DNA, Plant; D005998:Glycine; D009838:Oligodeoxyribonucleotides; D037121:Plant Lectins; D000097797:Glyphosate