1/12/2021 Difference Between DNA and RNA Extraction | Compare the Difference Between Similar Terms

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Difference Between Between DNA and RNA Extraction
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9, 2017 Posted by Samanthi (https://www.differencebetween.com/author/samanthi/)

Key Difference – DNA vs RNA Extraction

The study of DNA and RNA (https://www.differencebetween.com/difference-between-dna-and-rna/) are vital

aspects in order to understand the basic concepts of molecular biology
(https://www.differencebetween.com/difference-between-microbiology-and-vs-molecular-biology/),
biotechnology (https://www.differencebetween.com/difference-between-biotechnology-and-vs-biomedical-
engineering/), and genetics (https://www.differencebetween.com/difference-between-genetics-and-vs-
epigenetics/#Genetics). Extraction of pure DNA and RNA samples are necessary in order to carry out
experimental procedures during these studies. The key difference between DNA and RNA extraction is that
DNA extraction process purifies DNA while RNA extraction purifies RNA. DNA extraction process has
three different steps: cell lysis and catabolism (https://www.differencebetween.com/difference-between-
metabolism-and-vs-catabolism/)of membrane lipids and proteins, clumping of catabolites by concentrated salt
solution and the precipitation of DNA with ethanol (https://www.differencebetween.com/difference-between-
ethanol-and-vs-ethanoic-acid/). The three step procedure may consist of two optional steps. RNA purification
process consists of four different steps: addition of guanidium thiocyanate for cell lysis, protein denaturation

including ribonucleases, separation of RNA by the addition of chloroform and phenol

(https://www.differencebetween.com/difference-between-alcohols-and-vs-phenols/)and the washing of the
precipitate using ethanol.

CONTENTS

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1/12/2021 Difference Between DNA and RNA Extraction | Compare the Difference Between Similar Terms

CONTENTS

1. Overview and Key Difference

2. What is DNA Extraction
3. What is RNA Extraction
4. Similarities Between DNA and RNA Extraction
5. Side by Side Comparison – DNA vs RNA Extraction in Tabular Form
6. Summary

What is DNA Extraction?

DNA extraction is a physical and chemical process of that is used to purify DNA from a sample. DNA extraction
is an important aspect in the context of molecular biology and forensic sciences. The process is of three basic
steps. Initially, the cells of interest should be obtained. Next, cell lysis is facilitated in order to break the cell
membrane, which opens up the cell and exposes the cytoplasm
(https://www.differencebetween.com/difference-between-cytoplasm-and-vs-cytosol/)along with DNA.
Surfactants or other detergents can be utilized to lyse the lipids from the cell membrane while the proteins
present are catabolized by proteases. This is an optional step. Once the cell is lysed, clumping of the
catabolized molecules is facilitated by concentrated salt solutions. It is followed by centrifugation of the solution
which separates the debris clumps from DNA. In this stage, the differentiated DNA is mixed with the reagents
and salts used during the cell cycle.

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1/12/2021 Difference Between DNA and RNA Extraction | Compare the Difference Between Similar Terms

Figure 01: DNA extraction

In order to purify it further, the following steps could be used. One method is ethanol precipitation, which
involves mixing ice cold ethanol with the separated DNA sample. DNA is insoluble in alcohol and thereby
produce a pellet due to the aggregation of DNA molecules together. Sodium acetate is added in this process in
order to enhance the degree of precipitation by increasing the ionic strength. In addition to ethanol precipitation
process, phenol-chloroform extraction process could also be induced for this. In this method, phenol denatures
the proteins present in the sample. Once centrifuged, denatured proteins will remain in the organic phase while
the DNA molecules that are mixed with chloroform will be present in the aqueous phase. Chloroform will
remove the phenol residues. Once extraction is completed, DNA is kept dissolved in TE buffer or ultra pure
water.

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1/12/2021 Difference Between DNA and RNA Extraction | Compare the Difference Between Similar Terms

What is RNA Extraction?

RNA purification is a process by which RNA is purified from a biological sample. Due to the presence of
ribonuclease in the cells and tissues, this process is complicated. Ribonuclease enzyme has the ability to
degrade RNA rapidly. The chemical nature of ribonucleases is extremely stable, and it is hard to inactivate
them. Neutralizing the ribonucleases is an option. Since this enzyme is ubiquitous in the cells and tissues,
special techniques are developed for RNA extraction. Out of the many methods, the common method is
Guanidinium thiocynate-phenol-chloroform extraction.

Figure 02: RNA Extraction

The Guanidinium thiocynate-phenol-chloroform extraction method depends on centrifugation

(https://www.differencebetween.com/difference-between-filtration-and-vs-centrifugation/)and phase separation.
The mixture to be centrifuged consists of the aqueous sample and a water saturated solution that consist of
phenol and chloroform. Once centrifuged, the solution consists of an upper aqueous phase and a lower organic

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1/12/2021 Difference Between DNA and RNA Extraction | Compare the Difference Between Similar Terms

phase under neutral pH conditions (pH 7-8). RNA is present in the aqueous phase. The organic phase typically
consists of proteins dissolved in phenol and lipids dissolved in chloroform. A chaotropic agent (a molecule that
has the ability to break the hydrogen bonds between water molecules) is added; this is known as, guanidinium
thiocyanate. This agent has the ability to denature proteins which include ribonucleases that can degrade RNA
and is involved in cell lysis. It also separates rRNA from ribosomal proteins. The final step of RNA purification is
washing the precipitation of the aqueous phase with ethanol. RNA could also be purified using liquid nitrogen.

What are the Similarities Between DNA and RNA Extraction?

Both extraction processes utilize harmful chemicals such as phenol and chloroform.
Centrifugation is an essential technique for both processes.
Ethanol is used to wash the precipitate and to obtain purified DNA or RNA.

What is the Difference Between DNA and RNA Extraction?

DNA vs RNA Extraction

DNA extraction is a process which extracts DNA from an RNA extraction is a process which
organism or sample. extracts RNA from a sample.
Steps

The DNA extraction process is composed of three The RNA extraction process is composed
different steps with two optional steps. of four different steps.
Reagents

Surfactants, proteases (optional), alcohol, chloroform, Guanidium thiocyanate, chloroform,

phenol, sodium acetate are used for DNA extraction. phenol, ethanol are used for RNA
extraction.

Summary – DNA vs RNA Extraction

DNA and RNA extraction are vital aspects of the experimental procedures for the study of molecular biology,
genetics, and biotechnology. Both processes involve similar reagents, but RNA extraction utilizes a special
reagent known as Guanidium thiocyanate which diminishes the activity of ribonucleases. This is the main

difference between DNA and RNA extraction.

Download PDF Version of DNA vs RNA Extraction

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download PDF version here Difference Between DNA and RNA Extraction
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Extraction.pdf)

Reference:

1. Chomczynski, P, and N Sacchi. “Single-Step method of RNA isolation by acid guanidinium thiocyanate-
Phenol-Chloroform extraction.” Analytical biochemistry., U.S. National Library of Medicine, Apr. 1987, Available
here (https://www.ncbi.nlm.nih.gov/pubmed/2440339). Accessed 31 Aug. 2017
2. Hindawi. “DNA, RNA, and Protein Extraction: The Past and The Present.” BioMed Research International,
Hindawi, 30 Nov. 2009, Available here (http://www.hindawi.com/journals/bmri/2009/574398/). Accessed 31
Aug. 2017.

Image Courtesy:

1. “DNA extracted from avocado” By Mike Seyfang – originally posted to Flickr as success (CC BY 2.0)
(https://creativecommons.org/licenses/by/2.0) via Commons Wikimedia
(https://commons.wikimedia.org/w/index.php?curid=5162122)
2. “PhOH-CHCl3 extraction” By Squidonius (talk) – Own work (Original text: self-made) (Public Domain) via
Commons Wikimedia (https://commons.wikimedia.org/w/index.php?curid=13395851)

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About the Author: Samanthi

Dr.Samanthi Udayangani holds a B.Sc. Degree in Plant Science, M.Sc. in Molecular and
Applied Microbiology, and PhD in Applied Microbiology. Her research interests include Bio-
fertilizers, Plant-Microbe Interactions, Molecular Microbiology, Soil Fungi, and Fungal Ecology.

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