Isolate And Prepare Restriction Digest For The Thrombin Activatable Fibrinolysis Inhibition

1703 Words Aug 14th, 2015 7 Pages
GOALS
The aims of this lab report are to isolate and prepare restriction digest for the thrombin activatable fibrinolysis inhibition (TAFI) cloned to pcDNA.4A plasmid vector twice before introducing the site direct mutagenesis (SDM) and after the SDM. Indeed, SDM principle including bacterial transformation and sequencing principle will explained here. As well as, this lab report elucidates the RNA interference and its uses as a research tool by relative semi-quantifying of the phosphatase and tensin homologue (PTEN) and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression after they isolated from human embryonic kidney HEK 293 cells that either untreated or treated with siPTEN, siGAPDH or negative control siRNA. Lastly, an overview of flow cytometry including its components, principles, samples preparations, and data analysis.
BACKGROUND
Genetic engineering is the ability of researcher to manipulate the DNA at molecular level, which also refers to as recombinant DNA technology. This technology helps researchers to construct a novel DNA by recombining DNA species from different sources. Interestingly, most of the DNA knowledge elucidate from bacteria and bacteriophages. Indeed, most of the recombinant DNA technology tools such as plasmid, restriction endonucleases enzymes, DNA and RNA polymerases, and DNA ligases obtained from bacteria and bacteriophages. Endonucleases (also known as restriction enzymes) are biotechnology tools that researchers uses as scissors to…

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