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How is DNA collected?
It is also called "miniprep method". there certain other methods but i know about the miniprep method.It involves following basic steps:1. source to get the DNA like some samples2. increasing its quantity by PCR3. isolating the DNA out of unwanted substance into the pure form1. first the DNA containing solution is kept at the temp of 4 degree celsius so that all the enzymes get inactivated.2. it is treated with an anionic detergent so that the DNA makes a complex with it.3. the whole solution is treated with hot beta-merceptoethanol so that all the protein present gets denatured.4. it is then added with phenol:chloroform in the 24:1 ratio so that we get 2 layers.....a) upper organic layer which contains the nucleic acid i.e.DNA and RNAb) lower aqueous layer with cell debris and other unwanted material.5. the upper layer is decanted in another test tube6, now RNAse is added to remove RNA7. now NaOH is added to remove an anionic detergent.8. now ethanol is added to precipitate out the DNA in the form of smooling structure.
Function of GTE buffer solution?
GTE stands for Glucose-Tris-EDTA.Glucose is used to maintain osmolarity: 50mM (millimolar) glucose prevents premature cell lysis, which can cause lower DNA yields to due to aggregation and degradation. The other components of the buffer also contribute to the osmolarity of the solution, but glucose, being a non-electrolyte, is a good choice because it does not interfere with the solution's buffer properties. The Tris is used to buffer whatever you're adding this to at pH 7.9. EDTA binds divalent cations like Ca2+ and Mg2+, thereby weakening the cell envelope of cells in the mixture. This is typically used for miniprep/DNA purification, when you have to lyse the cell and get internal cell components out into the solution.
