Hi Could anyone please explain to me the principle behind the RNA isolation experiment.

Hi

Could anyone please explain to me the principle behind the RNA isolation experiment.

Expert Solution

RNA is a polymeric molecule made up of ribonucleotides. Ribonucleotides consists of ribose sugars and nitrogen bases known as adenine, uracil, cytosine and guanine. RNA is single stranded molecule found in living cells and genetic material of viruses. So, it is needed to isolate RNA which carries a lot of genetic information of virus particle.

Principle

TRIzol reagent is used which can maintain the integrity of RNA while homogenization of tissue and also helps in disruption of the cell components. Centrifugation is done after adding chloroform which separates the solution into two phases aqueous and organic. RNA remains in the aqueous phase. Separate the aqueous phase and RNA is precipitated using isopropyl alcohol. Further DNA and proteins can be separated from the interphase and organic phase using ethanol and isopropyl alcohol respectively. Recovered RNA from aqueous phase then can be used in cloning, in- vitro translation or in northern blotting etc.

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gladiator answered 1 year ago

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