dna isolation by ctab method -role of ctab,chloroform,phenol,isoamylalcohol

2 ANSWERS

1. Guest20967953
   

   why we prefer ctab for genomic DNA isolation from plant?

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2. Guest18768121
   

   1. When DNA has to be isolated from cells, CTAB is used to facilitate in the lysis of cells so DNA can be released into the bulk of the solution, from where is it is isolated through further steps 2.Efficient extraction of cell extracts or solutions containing nucleic acid are most often performed with a series of phenol and phenol:chloroform extractions at a specific pH. Both phenol and chloroform cause proteins to become denatured and become soluble in the organic phase or interphase, while nucleic acids remain in the aqueous phase. After centrifugation, the aqueous phase containing nucleic acid is re-extracted with an equal volume of chloroform:isoamyl alcohol (1,3,4). This combination of extractions is thought to reduce the loss of RNA due to the formation of insoluble protein:RNA complexes at the interphase. Chloroform is mixed with phenol to increase the efficiency of nucleic acid extractions by reducing losses of RNA at the interphase. The increased efficiency is due to chloroform's ability to denature proteins and aid in the removal of lipids, thus improving separation of nucleic acid into the aqueous phase. Phase separation is also enhanced, which assists in the removal of the aqueous phase with minimal cross contamination from the organic phase. Often isoamyl alcohol is added to phenol:chloroform to reduce foaming

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