DNA Cloning - Polymerase Chain Reaction (PCR)

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The body is a very interesting instrument developed by The Most High, we can't ignore the fact that we were created by a higher power; trust me you don't want to be created out of a big bang without no one watching.

In today's article we are going to be talking about, DNA cloning.

DNA stands for deoxyribonucleic acid, and it is a macro-molecule, that is very long; it is part of our body and it is responsible for transferring genetic information, in all life forms. The DNA is the main component of chromosomes, they are constructed of two nucleotide strands, which are coiled around each other, arranged similarly to a ladder.

The biology technique, which makes a lot of similar copies of a strand of DNA, such as a gene is known as DNA cloning. A plasmid is used, to insert a target gene in a typical cloning experiment.

The plasmid which is a circular piece of DNA is small and it is different from a cell's chromosomal DNA. Bacteria cells and some eukaryotes are the places where plasmid are extracted from. Plasmid genes give bacteria generic advantages, such as resistance to an antibiotic.

The wide range of lengths of plasmids goes from one thousand DNA base pairs to hundreds of thousands of base pairs. In the cloning process, through a technique called transformation, the plasmid is introduced into bacteria and antibiotics are used to select the bacteria which carry the plasmid. In many genetic engineering approaches to biotechnology, cloning is often the starting point.

In genetic engineering, large amounts of DNA are needed; therefore, multiple copies of DNA are made using various techniques. Polymerase chain reaction (PCR) is one of the techniques used to clone DNA and make several copies of a gene. There are of course steps to cloning DNA, you must first isolate or cut the DNA from its source, and then paste that target DNA onto a vector, which can then replicate itself.

Using restriction enzymes scientists are able to cut the chosen piece of DNA, DNA litigation is then used to paste the target DNA into a vector or plasmid. Through transformation, then the vector is introduced into a host cell, which is often yeast or bacteria. Once the copies are made the vector DNA is isolated and purified, from the host cell's DNA.

When PCR is used to make copies of DNA, the experimenter should be aware of the key ingredients needed for a PCR reaction to be successful. Primers, template DNA, Nucleotide and Taq polymerase are the key ingredients in a PCR reaction.

A tube is used to assemble the ingredients and co-factors needed by the enzyme, then a series of heating and cooling cycles, take place, for the DNA to be synthesized.

Denaturation is the first step in PCR, and what happens during this step is to simply, heat up the reaction to 96 degrees Celsius, to separate or denature the DNA strands. Once a single-strand template of DNA is ready, then the next step can take place.

During the next step, the reaction is cooled down to 55 – 65 degree Celsius, so that the primers can bind to the complementary sequences of the template DNA; this process is known as annealing. The temperatures are raised again in the reaction, to make Taq polymerase extend the primers, to form new strands of DNA. This cycle is repeated until enough copies of the target DNA is created.

Thank you for reading this article!!!

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Hernando Cadet

Hi every one, I obtained a bachelor's degree in Bioinformatics back in 2006, from Claflin University, after I received my bachelor's degree, I gained full time employment as a software engineer at a Video Relay Service company, maintaining databases and developing software for a new developed device called the VPAD.

I worked at that company for two years, then I became a web developer, and worked for a magazine for three years. After that job, I worked as a Drupal web developer, as a subcontractor for the NIH, for a year and then left the job to go back to school.

Hernando Cadet Hi every one, I obtained a bachelor's degree in Bioinformatics back in 2006, from Claflin University, after I received my bachelor's degree, I gained full time employment as a software engineer at a Video Relay Service company, maintaining databases and developing software for a new developed device called the VPAD.

I worked at that company for two years, then I became a web developer, and worked for a magazine for three years. After that job, I worked as a Drupal web developer, as a subcontractor for the NIH, for a year and then left the job to go back to school.