Creative Biogene has gathered a number of senior experts to provide the molecular characterization testing service to clients with stable quality and reasonable price. Creative Biogene is based on the frontiers of transgenic plant service with complete transformation system and years of service experience. And we have mature screening and testing technology for transgenic plants.
The molecular characterization testing of transgenic plants is an important step to determine whether the target gene has been transformed into the plants successfully. The molecular characterization testing of genetically modified organisms is of great importance. It is commonly applied to the safety assessment of genetically modified organism, laying the foundation for the subsequent monitor of genetically modified crops.
Molecular characterization testing service can be carried out from various levels (DNA, RNA and protein), our services include:
1. DNA Level
PCR screening is one of the most widely used methods because of the high efficiency, simple operation and low cost. PCR is an enzymatic reaction using DNA polymerase to amplify the target fragment. When it is used for analyzing transgenic plants, a band in specific size will be in the gel if the target gene is transformed into the plants successfully.
Southern Blotting is an assay utilizing labeled probes to specifically hybridize with the target DNA, which can be used for analyzing the integration of foreign genes on plant chromosomes and the stability of foreign genes in transgenic offspring. The southern blotting is used in molecular characterization of transgenes to determine the existence and copy numbers of the introduced gene into the GMO genome. It has been extensively used in the screening of transgenic plants of various crops such as rice, wheat, corn, soybean, rapeseed, peach with high accuracy and specificity.
2. RNA Level
RT-PCR is a technique that combines reverse transcription (RT) of RNA with polymerase chain amplification (PCR) of cDNA. In the transgenic plants, RT-PCR is used to detect whether the foreign gene is expressed at the mRNA level by electrophoresis detection. RT-PCR can detect low-abundance mRNA in transgenic plants, especially in the case of target genes is integrated in a single copy.
Northern blotting is a method to investigate gene expression by detecting specific RNA through electrophoresis and hybridization probe. It can be used to analyze the size and level of target RNA in transgenic plants. It is also known whether there is a relatively large amount of gene expression at the RNA level. Northern blotting is a significant means to study the expression and regulation of a foreign gene in transgenic plants such as potato, rice, tobacco.
Real-Time PCR is a technology to perform a total analysis or relative quantification. The key feature is that the entire PCR process is monitored by the fluorescence signal accumulation. It is useful to detect the relative expression of mRNA in transgenic plants compared to the control. It has the features of high sensitivity, accuracy and a high degree of automation to screen transgenic plants.
RNA Fluorescent In Situ Hybridization (FISH) is a widely-applicable histology technique that utilizes a nucleic-acid based probe to localize to RNA sequences of interest, and allows for visualization of RNA expression in cells or tissues. This method can be used in transgenic plants to detect and locate the presence and position of specific RNA sequences in the plant cells, providing insights into gene expression and function. RNA FISH assay as a powerful tool for studying the spatial-temporal regulation of gene expression in plant cells and tissues.
3. Protein Level
Western blotting is an extensively method to detect the expression of a specific protein. This method is used for qualitative result analysis of target proteins in GM plant samples. Based on the testing results, it is possible to know whether the target protein in transgenic plants is expressed, the concentration, and the approximate molecular weight. It has been widely used to detect protein expression of transgenic plants such as tobacco, Chinese wolfberry and poplar.
ELISA is a qualitative and quantitative screening method, which can be used to determine antigens or antibodies. It gives information about the quantity of protein in transgenic plant samples with high sensitivity. This method has been applied to the screening of various transformed plants such as cotton, pepper, rice, tobacco, and tomato.