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BTEC APPLIED SCIENCE: UNIT 11 - Learning Aim D

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Genetic Engineering. At a grade Distinction.

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Unit 11: Genetics and Genetic Engineering Learning Aim D: Explore basic DNA
techniques and the use of genetic engineering technologies.


Index.
 Introduction,

 Extract, Amplify and Separate DNA,

 Use of genetic Engineering Technologies in Industry and Medicine,

 Analysis of the uses of Genetic Engineering Technologies in Industry and
Medicine,

 Evaluate possible future uses of Genetic Engineering Technologies,



Introduction.

This report will show how to extract, amplify and separate DNA, Explain the use of genetic
engineering technologies in industry and medicine. Analyse the uses of genetic engineering
technologies in industry and medicine and Finally Evaluate possible future uses of genetic
engineering technologies.

Genetic Engineering.

Identifying the segment of DNA that contains the appropriate gene is the first step in genetic
engineering. Following that, the gene is extracted and incorporated into a bacterial plasmid. After that,
the plasmid is inserted into the host cell. These transformed cells can multiply and create a GM
organism.

DNA extraction of a banana method.

Step 1: Place the banana into a zip-closure bag and remove most of the air before you seal the bag,

Step 2: Mash the banana through the bag in your hand. Do not hit against the table as this might
damage the DNA,

Step 3: Add 2 tablespoons of the DNA extracting solution,

Step 4: Continue mixing and mashing the bag in your hand,

Step 5: Place a piece of gauze over opening of the cup, securing it with a rubber band,

Step 6: Carefully pour the banana mixture into the cup making sure to catch the solids with the
gauze,
1

, Step 7: Take a drop or spoonful of the liquid in the cup and place in the test tube,

Step 8: Add a drop or spoonful of the alcohol to the test tube. Take care not to tilt or tip the test tube;
do not mix the two liquids,

Step 9 : Observe the line between the banana mixture and the alcohol.

The cells of the banana are burst open by the dishwashing liquid therefore releasing the DNA. The
salt used ensures that the proteins in the cell are kept separate from the DNA. Furthermore Insoluble
molecules clump together and become apparent as they are unable to be dissolved. Since DNA
strands are not soluble in alcohol, they clump together and become visible to the naked eye.

A sample's cells are separated from one another, generally by physical techniques such as
compressing, and then placed in a salt solution. The negatively charged phosphate groups that run
along the backbone of DNA are protected by the positively charged sodium ions in the salt. The lipids
in the cell membrane and nuclei are then broken down using a detergent. These membranous
structures release DNA.

To obtain a clean sample of DNA, as much cellular debris as possible must be removed. This can be
accomplished in a number of ways. To breakdown DNA-associated proteins and other cellular
proteins, a protease (protein enzyme) is added. Filtering the sample can also help eliminate some of
the cellular debris.

Finally, the DNA sample is carefully added with ice-cold alcohol (either ethanol or isopropanol). Water
makes DNA soluble, but salt and alcohol make it insoluble. A precipitate can be seen and spooled out
by gently swirling the alcohol layer with a sterile pipette. A stringy, white precipitate may form if there
is a lot of DNA present.

The DNA sample can now be purified even more. DNA can be extracted and used for molecular
analyses such as PCR, electrophoresis, sequencing, fingerprinting, and cloning once it has been
resuspended in a slightly alkaline buffer.

PCR.

PCR stands for Polymerase chain reaction. It is the method of rapidly multiplying a single DNA
molecule into millions of copies. Denaturation is the process of heating double-stranded DNA to
separate the strands. The primers bind to flanking regions of the target DNA during the annealing
process. The DNA polymerase extends the end of each primer along the template strands in the final
stage, extension. PCR can be used for cloning of a single sequence.

Denaturation, or separation, of the two strands of the DNA molecule is the first step. This is performed
by heating the starting material to around 95 ° C. Each strand serves as a blueprint for creating a new
strand. The temperature is decreased to around 55 °C in the second phase so that the primers can
anneal to the template. The temperature is raised to around 72 °C (162 °F) in the third phase, and the
DNA polymerase begins adding nucleotides to the annealed primers' ends. The temperature is raised

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