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 In addition, RFLP from homozygous organism (AA), heterozygous organism (AS), and homozygous organism (SS) is produced in various lengths by Mst II enzyme.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 So, Mst II is not able to recognize this mutated sequence (CCTGTGG). As we know, when adenine nucleotide in the sequence CCTGAGG mutates into thymine nucleotide, the sickle-cell disease appears. Mst II will not recognize and will not cut, if DNA has a mutation from sickle cell disease. Mst II cuts non-mutated (normal) beta globin DNA at a particular site, which is CCTNAGG (N is any base). The restriction enzyme called Mst II is used in the identification of sickle-cell disease. Generally, restriction enzymes, binding to DNA, slide along a DNA until they recognize specific sequences of nucleotides, where they start to perform their work, i.e. Restriction enzymes are the enzymes, which cut at specific sequence of nucleotides called restriction sites. Finally, RFLPs can be needed to identify genetic disease as well as disease carrier. Also, RFLPs are used to avoid inbreeding by the identification of heterogeneous parents. We use RFLPs in forensic labs to determine a criminal or we can use RFLPs to determine fatherhood. Restriction fragment length polymorphisms (RFLPs) are fragments of various sizes created by polymorphic region on DNA, when restriction enzymes cut (digest) DNA. Also, some radioactive elements can be absorbed by human’s body becoming incorporated in bones, and it is actually, very hard to eliminate an absorbed radioactive element from the body. For example, even a small amount can cause cancer (sometimes affecting DNA). We do not use radioactive probes to detect DNA in Southern Blot Analysis because radioactive isotopes (radioactive elements) can cause very dangerous effects on human’s organism when they are not handled properly. Generally, to perform the technique, separated DNA fragments obtained by electrophoresis are transferred to a filter membrane and then, detection by probe hybridization is made. The Southern blot analysis is a technique that allows determining specific DNA sequence in a sample of DNA. Southern Blot Analysis - Sickle Cell Diagnosis Lab
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