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  • PROCUREMENT OF DNA AND SEPARATION OF DNA FRAGMENTS

    images16A molecular biology laboratory come from clinical different types of samples, each with its own peculiarities to the analysis and obtain his DNA. “A priori” two different situations arise. If the aim is to know the physical location of the DNA / RNA of interest in a histological section shall be undertake studies in situ, whereas if it is not necessary to know that location is sufficient to obtain cellular DNA in solution. The DNA in solution is a mixture of mitochondrial DNA, eukaryotic nuclear and viral / bacterial if present in the sample virus / bacteria.

    Focusing on the extraction of DNA in solution are different methods of analysis: differential lysis, lysis neutral … After the lysis, the DNA solution obtained is not always suitable for use in subsequent amplification reactions, restriction, labeling or sequencing, so that one side should separate the DNA from other macromolecules that accompany it, preferably proteins, a process that usually performed using solvents organic (phenol and chloroform) and secondly to separate the DNA of those small molecules with inhibitory activity present in the DNA solution. The once clean DNA protein and / or small molecules can be used directly, although in most situations is to concentrate by ultrafiltration or precipitation with alcohol.

    The rating system is the concentration of DNA which were based on treatment and therefore the degree of purity of DNA obtained. Can be used from highly sensitive and accurate methods such as spectrophotometry and fluorometry, to approximate methods such as comparison the degree of fluorescence with respect to a control of known concentration.

    Published on October 27, 2012 · Filed under: DNA; Tagged as: , , , ,
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