Isolated Dna Products Amplified Via Polymerase Chain Reaction and Cloned
Isolated DNA Products Amplified
Via Polymerase Chain Reaction and Cloned
Biotechnology: DNA
WPUNJ
December, 2012
Abstract
Isolated DNA from mouse, plants, and plasmid DNA were used for Polymerase Chain Reaction (PCR) for DNA amplification. The purpose of this experiment was to study the success rate or optimization of PCR of DNA, using both manual and kit methods. This set of experiments gives an insight to the relative difficulties associated with the optimization of a variety of DNA/primer paired samples. Through this experimentation, researchers were successfully able to assess the issues and difficulties in both the manual and kit optimizations that were performed. Through this report, step by step methods used to achieve viable PCR product are discussed, along with different methods of trouble shooting and experiences through out the experimentation. The PCR product’s amplified DNA was then used for insertion into a plasmid vector system, which too resulted in various range of consequences and difficulties. Hence, the success rate and difficulties are viewed and analyzed through out this paper as experimenters try to achieve maximal desired PCR products associated with each protocol used.
Introduction
Polymerase Chain Reaction (PCR) is a technique used in molecular biology and biotechnology, to amplify a small amount of DNA in a reaction. This simple and inexpensive technique is a fundamental tool that focuses on the segment of given DNA and copies it over. Today, PCR is used in every day laboratories to study diseases, identify bacteria or viral DNA, and trace criminals.
To conduct a PCR reaction, a few critical elements are needed, such as: polymerase, primers, nucleotides, buffers, and the most crucial, the template that needs to be amplified. DNA polymerase, acquired from Thermis aquaticus, is the most important enzyme in this reaction, as it adds complementary bases to the template and binds to the region that needs to
Via Polymerase Chain Reaction and Cloned
Biotechnology: DNA
WPUNJ
December, 2012
Abstract
Isolated DNA from mouse, plants, and plasmid DNA were used for Polymerase Chain Reaction (PCR) for DNA amplification. The purpose of this experiment was to study the success rate or optimization of PCR of DNA, using both manual and kit methods. This set of experiments gives an insight to the relative difficulties associated with the optimization of a variety of DNA/primer paired samples. Through this experimentation, researchers were successfully able to assess the issues and difficulties in both the manual and kit optimizations that were performed. Through this report, step by step methods used to achieve viable PCR product are discussed, along with different methods of trouble shooting and experiences through out the experimentation. The PCR product’s amplified DNA was then used for insertion into a plasmid vector system, which too resulted in various range of consequences and difficulties. Hence, the success rate and difficulties are viewed and analyzed through out this paper as experimenters try to achieve maximal desired PCR products associated with each protocol used.
Introduction
Polymerase Chain Reaction (PCR) is a technique used in molecular biology and biotechnology, to amplify a small amount of DNA in a reaction. This simple and inexpensive technique is a fundamental tool that focuses on the segment of given DNA and copies it over. Today, PCR is used in every day laboratories to study diseases, identify bacteria or viral DNA, and trace criminals.
To conduct a PCR reaction, a few critical elements are needed, such as: polymerase, primers, nucleotides, buffers, and the most crucial, the template that needs to be amplified. DNA polymerase, acquired from Thermis aquaticus, is the most important enzyme in this reaction, as it adds complementary bases to the template and binds to the region that needs to
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