DNA Extraction: Learn to Extract DNA

Learn to Extract DNA from Plants as well as from Blood, Become an DNA Extraction Expert

4.29 (7 reviews)
Udemy
platform
English
language
Science
category
instructor
15
students
30 mins
content
May 2022
last update
$34.99
regular price

What you will learn

About different DNA Extraction Methods

How DNA Extraction is Performed

How DNA Extraction from Plants is different then from Blood

Introduction to Genome

Description

DNA extraction is a method to purify DNA by using physical and/or chemical methods from a sample separating DNA from cell membranes, proteins, and other cellular components. Friedrich Miescher in 1869 did DNA isolation for the first time.

The use of DNA isolation technique should lead to efficient extraction with good quantity and quality of DNA, which is pure and is devoid of contaminants, such as RNA and proteins. Manual methods as well as commercially available kits are used for DNA extraction. Various tissues including blood, body fluids, direct Fine needle aspiration cytology (FNAC) aspirate, formalin-fixed paraffin-embedded tissues, frozen tissue section, etc., can be used for DNA extraction.

DNA extraction involves lysing the cells and solubilizing DNA, which is followed by chemical or enzymatic methods to remove macromolecules, lipids, RNA, or proteins.

DNA extraction techniques include organic extraction (phenol–chloroform method), nonorganic method (salting out and proteinase K treatment), and adsorption method (silica–gel membrane).

Organic extraction:

This method is labor intensive and time consuming.

Cell lysis can be done using nonionic detergent (sodium dodecyl sulfate), Tris–Cl, and Ethylene diamine tetraacetic acid (EDTA), and this step is followed by removal of cell debris by centrifugation. Protease treatment is then used to denature proteins. Organic solvents such as chloroform, phenol, or a mixture of phenol and chloroform (phenol/chloroform/isoamyl alcohol ratio is 25:24:1) are used for denaturation and precipitation of proteins from nucleic acid solution, and denatured proteins are removed by centrifugation and wash steps. RNAse treatment is done for the removal of unwanted RNA. Precipitation with ice-cold ethanol is performed for concentrating DNA. Nucleic acid precipitate is formed, when there is moderate concentration of monovalent cations (salt). This precipitate can be recovered by centrifugation and is redissolved in TE buffer or double-distilled water.

Other methods include silica-based technology (DNA absorbs to silica beads/particles at a specific pH in presence of specific salts), magnetic separation (DNA binds reversibly to magnetic beads, which are coated with DNA-binding antibody), anion exchange technology, salting out, and cesium chloride density gradients.

Assessing the quality and yield of DNA: The quality and yield of DNA are assessed by spectrophotometry or by gel electrophoresis. Spectrophotometry involves estimation of the DNA concentration by measuring the amount of light absorbed by the sample at specific wavelengths. Absorption peak for nucleic acids is at ~260 nm. The A260/A280 ratio is ~1.8 for dsDNA. A ration of less than 1.7 indicates protein contamination.

Content

Introduction to DNA Extraction

Introduction to Course and DNA Extraction Reasoning
Introduction to making your own DNA Glass Necklace
Protocol of Making your own DNA Necklace

DNA Extraction from Plant

DNA Extraction Method Types along with Reagents and Materials Requirement
Procedure for Extracting DNA from Plants
Agrose Gel Quality Control (QC)

DNA Extraction from Blood

Introduction to DNA Extraction from Blood
DNA Extraction from Blood and it,s Protocol

Genome

Introduction to Genome
Article on Genome
Central Dogma

About DNA Extraction (Reading)

Importance, Discovery, Application

Screenshots

DNA Extraction: Learn to Extract DNA - Screenshot_01DNA Extraction: Learn to Extract DNA - Screenshot_02DNA Extraction: Learn to Extract DNA - Screenshot_03DNA Extraction: Learn to Extract DNA - Screenshot_04

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4648946
udemy ID
4/19/2022
course created date
7/17/2022
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