The modified technique will allow rapid analysis for large sample numbers to overcome the drawbacks of conventional methods.Ģ.1. PCR methods using transgene as allowing rapid detection of transgenic DNA, screening method is based on the amplification of the extracted DNA which is suitable to target primer sequence according their difference in length which is called amplification using complementary primers, this method can lead to undesirable background of unwanted and non-specific amplifications. In spite of its accurate results, it is labouring and time consuming method especially during assaying many samples and can consume important portions of the genomic DNA. In this report, seeds (T1 generation) will be used of N70756 strains obtained from Arabidopsis plants transformed with an Agrobacterium-mediated transformation, after extracting the genomic DNA from plant PCR based method will be used and Southern blot for detection, analysis and screening transgenic plants, Southern blot analysis is the most popular method for identification of transgenic DNA and also the copy number. Methods and analysis procedures used for getting transgenic Arabidopsis plants by transformations with Agrobacterium results in generation of various number of DNA copies at different locations on the plant chromosomes, these copies can affect the stability and the level of the expression of the gene so the identification of the transgenes should be conducted for familiar analysis of transgenic expression levels which should be unique procedures. Arabidopsis species and then cloned using plant vector transformation in the presence of a suitable marker which is antibiotic resistance for best selection of successfully transformed plant which is grown in the media with antibiotic after transformation. Īgrobacterium is a gram-negative bacteria used for gene transfer as it has the ability to transfer DNA between other plants, fungi and itself so it is a crucial tool for plant genetic engineering for improvement of the plant by transformation, the gene is recombined with the plant e.g. PCR give information about DNA of Arabidopsis, applications of theĪpproach to Arabidopsis plants after transformation with an Agrobacterium-mediated transformationĪrabidopsis is a plant from the family Cruciferae or Brassicaceae, it is one of the flowering plants related to white and black mustard this genus of plant is ideal for studying plant biology as the first complete sequence studied was the seeds of this plant for getting the complete genome. Of products of PCR result can be considered as a good estimation of transgenicĭNA, during comparison to southern blot analysis, the results obtained by the Plants from transgenic and non-transgenic cell lines.
The methods were specific and reproducible for many different Modifications in the methods as ligating PCR technique developed forĪmplification of the unknown DNA after extracting it to determine its Possible because the length of the given primers, there are slightly For identification of transgenic DNA of Arabidopsis plant material, the followed methods are transformation and growing of seedsĪfter bleaching them f or sterilization shouldīe performed then using polymerase chain reaction (P CR) for amplification the gene before making gelĮlectrophoresis to be ready for southern blot analysis, this method is
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