biochem-lab-report-3 CHEM 378.LB|very helpful

CHEM 378.LBBryant Peng Lab Partner Richmond Lam Biochemistry Lab Friday 4-8 Biochemistry Lab Report 3 Abstract: In these experiments we focused on the biochemical manipulation of genes with enzymes by using the various recombinant DNA technique and technology that are available to us. Some techniques we learned was the cutting of vectors with restriction endonucleases allowing us to map the restriction sites in our vectors. The restriction fragments formed by these cuts can be measured with an agarose gel electrophoresis to determine overall band size. Another manipulation of DNA we learned was the joining of cells with a ligation mixture composed of DNA fragments, ATP buffer, T4 DNA ligase, and water. Over time in a sticky end ligation there will always be one band representing our ligation fragment. An introduction of recombinant DNA into a host cell transforming the original host into something new. Because of the inefficiency in transformation antibiotic resistance is put in with the vector to identify the transformed host cell. In our experiment however the transformation turned out poorly and majority of the host cell died out instead of forming their own colonies and prosper. A major manipulation of DNA is PCR in which we amplify a set of gene for analysis which we do with the human cheek cell to test for the type Alu insertion in our gene. From our PCR reaction we learned that my partner in this experiment contained a heterozygous insertion of Alu gene. The many technique and technology available for us to manipulate genes biochemically were put into practice throughout this experiment. Introduction: In these set of experiment we utilized various different recombinant DNA techniques to biochemically manipulate genes. These techniques are important because they allow us to study an individual set of genes by creating a DNA molecule containing our gene of interest. In order to achieve such an outcome a vector is necessary for our experiment. A vector is a replicon, a piece of DNA able to function as a chromosome in vivo, which can carry our gene of interest in its DNA. A vector is expected to have the following properties: able to function in various host organism, multiple selection markers, and a high-level of expression for our gene of interest. A common vector perfect for recombinant DNA technique would be the plasmid. A plasmid is a small circular double-stranded extrachromosomal DNA which consist of an origin of replication (ORI), an antibiotic resistant gene, and a multi cloning site (MCS). A replication of origin (ORI) is a unique genomic sequence in DNA that initiates replication of the gene. An antibiotic resistant gene in a plasmid allows for selection later on in the experiment. A multi cloning site (MCS) is a short segment of DNA with m