The detection of a transgenic soybean biochip using gold label silver stain technology
| Autor: | Ke Han, Zhongliang Deng, Yuetao Ge, Qike Cao |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Streptavidin
Silver Staining Clinical Biochemistry Pharmaceutical Science Biochemistry Sensitivity and Specificity law.invention Silver stain chemistry.chemical_compound Biotin law Drug Discovery Biochip Molecular Biology Polymerase chain reaction Oligonucleotide Array Sequence Analysis Chromatography Chemistry fungi Organic Chemistry food and beverages Plants Genetically Modified Molecular biology Colloidal gold Nucleic acid Molecular Medicine Nanoparticles Gold Soybeans DNA microarray |
| Zdroj: | Bioorganicmedicinal chemistry letters. 21(22) |
| ISSN: | 1464-3405 |
| Popis: | A method for the rapid detection of transgenic soybean crops based on a combination of gene chip and "gold label silver stain" (GLSS) technologies has been established. To ensure the specificity of this method, the CaMV35S promoter and Nos terminator were selected as probes because they are both exogenous genes that are specific to transgenic soybean plants. The addition of biotin-modified dUTPs to a polymerase chain reaction (PCR) system can produce amplified nucleic acid segments containing biotin. These labeled PCR products then hybridize with specific probes on the chip and are subsequently bound by streptavidin-modified gold nanoparticles (GNPs). Due to the catalytic nature of the GNPs, silver staining can be used to visualize the hybridized probes, which appear as signals in varying shades of gray. The intensity value of the gray signals can be obtained using a general scanner. Silver staining for 10 min was determined to produce the optimal signal-to-noise ratio. In addition, this method was shown to be highly specific and had a detection sensitivity of 288.57 pg/μL. |
| Databáze: | OpenAIRE |
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