This document describes the process of extracting DNA from plant cells. It involves crushing plant material like strawberries or kiwi to break open cell walls and membranes. A saline-soap solution is added to further rupture cells and release DNA. Proteins and lipids are then filtered out, leaving the DNA in solution. DNA is precipitated by adding alcohol, allowing it to be collected and observed. The goal is to perform a simple, low-cost DNA extraction to understand the process at a molecular level.
This document describes the process of extracting DNA from plant cells. It involves crushing plant material like strawberries or kiwi to break open cell walls and membranes. A saline-soap solution is added to further rupture cells and release DNA. Proteins and lipids are then filtered out, leaving the DNA in solution. DNA is precipitated by adding alcohol, allowing it to be collected and observed. The goal is to perform a simple, low-cost DNA extraction to understand the process at a molecular level.
Original Description:
Extracción de ADN detallado con procesos que se pueden llevar en el laboratorio
This document describes the process of extracting DNA from plant cells. It involves crushing plant material like strawberries or kiwi to break open cell walls and membranes. A saline-soap solution is added to further rupture cells and release DNA. Proteins and lipids are then filtered out, leaving the DNA in solution. DNA is precipitated by adding alcohol, allowing it to be collected and observed. The goal is to perform a simple, low-cost DNA extraction to understand the process at a molecular level.
This document describes the process of extracting DNA from plant cells. It involves crushing plant material like strawberries or kiwi to break open cell walls and membranes. A saline-soap solution is added to further rupture cells and release DNA. Proteins and lipids are then filtered out, leaving the DNA in solution. DNA is precipitated by adding alcohol, allowing it to be collected and observed. The goal is to perform a simple, low-cost DNA extraction to understand the process at a molecular level.
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Deoxyribonucleic acid or DNA is the genetic material of living beings.
The extraction of DNA is
done with the purpose of doing genetic, science and molecular studies. We can extract the DNA in the easy way with products and a little care.
Objectives of DNA extraction
DNA consists of two chains of nucleotides, joined together forming a double helix. The skeleton has phosphate groups that are negatively charged and polar, which gives the DNA a negative net charge. Within the nucleus, this molecule is packed with proteins around it.
The objectives of this DNA extraction are:
Apply a simple technique to obtain DNA from plant cells.
Understand the process necessary for DNA extraction.
The isolation of DNA using the protocol with common salt and dishwasher is a simple, easy, fast, economical and non-polluting alternative. It is based on the physicochemical characteristics of the nucleic acid.
Steps for DNA extraction
The procedure to perform DNA isolation follows the following steps:
Step 1: preparation of saline-soap solution
Prepare a saline-soap solution composed of 100 ml of water (half a cup), 10 ml of dishwashing
detergent (1 tablespoon) or 20 ml of shampoo (without conditioner, two tablespoons) and 13 g of NaCl or kitchen salt (one tablespoon) ).
Step 2: preparing the fruits
If we use kiwi or banana, we peel and cut it into small cubes, then grind it in a mortar or liquefy it. If we use strawberries, we can place them in a bag with airtight seal and crush them.
Step 3: adding the salt and detergent solution to the fruit
In the mortar with the crushed fruit (or in the bag with the strawberries) pour the saline-soap solution and continue the process of breaking the fruits. Optionally, we can place the preparation in a water bath at 37ºC for 15 minutes to facilitate the extraction
Step 4: separation of solid material, proteins and lipids
The salty-soapy fruit juice is strained and filtered to remove the solids, proteins and lipids from the aqueous solution of the DNA.
We passed through a strainer or filter paper to remove the thickness of the fruit soup and then, through the gauze or cotton, to obtain 5 ml of the filtered liquid in a test tube or 50 ml in a jar. Step 5: precipitation of DNA by alcohol
Between the aqueous phase and the alcohol phase begins to observe the DNA fibers.
On the fruit juice, pour slowly through the walls of the ethanol or cold isopropanol test tube. Let rest a few minutes. A whitish, gelatinous layer should appear.
If we introduce a glass rod or a wooden stick, with circular motions we can recover the DNA wrapped in the rod.
Explanation of the stages of DNA extraction
Plants have three types of DNA: nuclear, mitochondrial and chloroplast. In addition, most plants have polyploidy, that is, the condition of some cells or organisms possessing more than one set of chromosomes.
Mature strawberries (Fragaria x ananassa) are an excellent material to extract DNA. They are easy to crush and contain enzymes, called pectinases and cellulases, that help break down cell walls. In addition, strawberries are octoploid (7 chromosomes x 8 = 56 chromosomes), that is, they have eight copies of each chromosome, which provides more DNA than banana or kiwi.
The kiwi has 29 chromosomes and is tetraploid. The banana or banana has 11 chromosomes and the majority is triploid. But you can also do this experiment with onions, lentils or other vegetable.
Rupture of tissues, walls and cell membranas
The detergent molecules help break down the cell membranes and release the DNA from the nucleus.
By friction with the mortar or by liquefying the material, the junctions between the cells and the cell wall are broken. This homogenization facilitates the lysis or rupture effect that helps to release the genetic material.
Soap or liquid detergent helps break down cell membranes, which are composed of lipids.
Elimination of proteins and lipids
Non-soluble components such as fibrous material and proteins are separated from DNA by filtration.
DNA precipitation
After the lipids and proteins are removed, the phosphate groups in the DNA are negatively charged and polar. DNA dissolves in aqueous solutions, but it is insoluble in alcohol. The addition of ethanol and high concentrations of sodium ions that bind to the phosphate groups, reduces the repulsive forces between the chains and allows the DNA to fold on itself making it insoluble. In this way it precipitates and forms a whitish filamentous mesh containing proteins and other materials.
