Tuesday, May 12, 2020

Polymerase Chain Reaction Essay - 1003 Words

One may view cloning as copying a living thing and producing multiple copies. People may think of cloning rabbits, sheep or humans. In the field of molecular biology, however cloning is viewed at a genetic molecular level, where a piece of DNA is copied on a large-scale by genetically copying tens to hundreds of thousands of identical DNA fragments. Researchers are developing new methods of cloning by using polymerase chain reaction (PCR). PCR was introduced in the 1980s and in recent years Kary Mullis won the Nobel Prize in Chemistry for his invention of PCR. Today, Scientists today are researching the various sub-fields of cloning, using PCR, in new ways using terminators, enzyme insertion, and types of cloning to produce high†¦show more content†¦This article also explain how polymerase chain reaction (PCR) is the most powerful amplification technology available for producing large quantities of DNA from a sample (Mullis et al. 2006). The scientists also tested thermo sta ble DNA polymerase and found Thermus aquaticus Polymerase (Taq) is the best for polymerase chain reaction. PCR is composes of three steps; denaturation, primer annealing, and polymerization. In the denaturation step, the target DNA is separated into two stands through heating, the hydrogen bonds between complementary bases, yielding single stands of DNA. In the annealing step, the temperature is decreased to anneal the primers. In the polymerization step, the template DNA is used by Taq polymerase to produce a complementary copy by extending the primers from their 3’ ends of the DNA. The development of thermo stable polymerases based on Taq, T4 DNA, and pfu polymerase, revolutionized PCR and converted it to a technique that can be used routinely in any lab. In respect to molecular biology, cloning has been the center for branching out new technologies in the topic to cloning. In Cloning and analysis of PCR-generated DNA fragments (Coasta et al. 1994), had explored the five main methods for cloning. They include, restriction enzyme site incorporation, T/A cloning, Uracil-DNA-gylcosylase cloning (UDG), ligase independent cloning, and blunt ended cloning. Each of the five many cloningShow MoreRelatedThe Reaction Of Polymerase Chain Reaction1823 Words   |  8 PagesPolymerase Chain Reaction: Polymerase chain reaction, also known as PCR, a technology that has made a tremendous impact on researchers, and has also affected many aspects of our everyday lives. The introduction of recombinant DNA technology has revolutionized the study of life as a tool for the biological sciences. Molecular cloning allowed the study of individual genes of living organisms; however there was dependence of obtaining a relatively large quantity of pure DNA. Scientists found it extremelyRead MorePolymerase Chain Reaction ( Pcr )1564 Words   |  7 PagesMBB/BIO 181 | Polymerase Chain Reaction (PCR) Name: Teresa Naval INTRODUCTION Scientifically, why is the study of Alu insertions interesting? (10 pts) Alu insertions are biological tools that can trace human history, evolution, and migration. In particular, Alu is often used to understand prehistory because all primates (including humans) with Alu insertions at specific locations can be traced to common ancestors [3]. Alu is sometimes referred to as a â€Å"jumping gene† because it copies itselfRead MoreDna Sequences Using Polymerase Chain Reaction1605 Words   |  7 PagesAmplification of 16S Ribosomal DNA Sequences using Polymerase Chain Reaction Edwina Abou Haidar, Houssam Al Koussa, Mary AbedAlAhad. Department of Biology, Lebanese American University, Byblos, Lebanon Abstract The 16s rRNA gene sequencing is a widely common amplicon sequencing method used to identify and compare bacteria in a given sample. This method is well established and allows to study phylogeny and taxonomy of complex microbiomes. In this study, an unknown sample of extracted microbialRead MoreEssay What is the Polymerase Chain Reaction (PCR) 1033 Words   |  5 PagesThe polymerase chain reaction or PCR for short can be used to create many copies of DNA. This allows the DNA to then be visualized using a dye like ethidium bromide after gel electrophoresis. The process has been refined over the years, however the basic steps are similar. The first is to denature dsDNA through heating to ~96  °C. This separates the two strands of DNA. The exact temperature to be used can be calculated with Tm = 4oC x (no. of G C) + 2oC x (no. of A T). Tm is the melting pointRead MoreLab Report : Bacillus Subtilus Using A Polymerase Chain Reaction1903 Words   |  8 Pages The purpose of this experiment was to amplify the ÃŽ ±-amylase gene in Bacillus subtilus using a polymerase chain reaction. Bacillus subtilus is a gram-positive bacterium that resides in soil and the gastrointestinal tract of animals and primarily uses aerobic respiration for survival (Yu et al., 2015). Due to it’s ability to take part in the fermentation process, B. subtilus can produce large amounts of the ÃŽ ±-amylase enzyme which is responsible for catalyzing the digestion of polysaccharides, suchRead MoreA Short Note On A Polymerase Chain Reaction ( Pcr ) Essay1264 Words   |  6 PagesMy longest ongoing project began in October 2014 under the guidance of Dr. Daniela Vergara, a postdoctoral researcher of the CGRI. Dr. Vergara guided another undergraduate student and me to troubleshoot a protocol for a Polymerase Chain Reaction (PCR). The objective of improving this protocol was to analyze phylogenetic relationships between local samples by amplifying and comparing a regi on of Tetrahydrocannabinol Acid (THCA) Synthase Gene. My colleague and I had applied for the Undergraduate ResearchRead MoreDna Barcoding Using Coi For Species Identification For Conservation1573 Words   |  7 PagesThe DNA Barcoding is becoming more popular in the present times due to its accuracy in the identification of different species. It has been approved to be more accurate than other taxonomic methods. The DNA Barcoding employs in the use of Polymerase Chain Reaction to magnify the COI gene. The amplified COI genes of the organisms are sequenced and compared with a known database of the organisms. The DNA Barcoding aids in understanding many characteristics of the species. These characteristics of theRead MoreEssay on Using PCR and Gel Electrophoresis to Determine Genotype583 Words   |  3 PagesGel Electrophoresis to Determine Genotype In certain situations, it is necessary to identify DNA retreived from a sample. When there is a small sample in need of identification, Polymerase Chain Reactions are used to multiply the DNA in the sample in to many identical samples. The DNA retrieved from the reaction can then be imported into an aparatus using gel electrophoresis to compare the sample of DNA to other samples. In our experiment we learned the how to replicate tiny samples of DNARead MoreAbstract. Taq Polymerase Is Essential In Polymerase Chain1446 Words   |  6 Pages ABSTRACT Taq Polymerase is essential in Polymerase Chain Reaction(PCR) experiments to obtain a PCR amplification of an unknown gene. The unknown gene is then ligated into a vector plasmid, which is placed in a bacterium Escherichia Coli to transform and multiply. Ultimately, identification and characterization of the unknown gene is done using electrophoreses and gel imaging. Cloning techniques such as the one performed have been used for many years to isolate genes from a variety of species.Read MoreA Segment Of The Will Die Slowly Gene From Drosophila Melanogaster1688 Words   |  7 PagesHarden’s lab at Simon Fraser University using random PCR primers. Attempts at purifying and characterizing this gene have been elusive. Here, we show methods for isolating, amplifying, and purifying the gene of interest for analysis. Using polymerase chain reaction to amplify the gene, it is then ligated into a pGEM-T Easy vector for TA cloning experiments. Transformation for further vector prolif eration is done on competent Escherichia coli cells. Subsequently, the vector is purified through plasmid

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