From that point on, new nucleotides are added to each of the original strands (A to T, C to G) until the result is two identical sequence copies of DNA. 3. How is DNA information used to synthesize polypeptides? A gene or protein is used to make polypeptides. In order to create this gene, transcription and translation must take place to create a protein from DNA.
Amir Ahemedin Ms.Buckley Genetics 11/06/15 Transformation of E.coli Lab Purpose The purpose of this lab is to genetically engineer the E.coli strain by introducing two genes, the green fluorescent protein gene (GFP) and the ampicillin resistant gene (AMP). Then observe whether or not the E.Coli strain would take up these genes and become fluorescent. Background Information In this lab, bacterial transformation was one of the three processes that occurred when genetic material is introduced to a bacterial cell. Bacterial transformation is important because it allows for the cloning and movement of DNA between strains. This transformation usually occurs within plasmids, which are closed circular molecules made up of double stranded DNA.
Title: Genetic pGLO Transformation Introduction: Genetic transformation is a transformation that involves a change in genes. Transformation is the process by which the genetic material carried by a cell is altered by the incorporation of foreign exogenous DNA into its genome. In this lab, we used a procedure called heat shock, accompanied by a bacterial plasmid vector, to transform bacteria with a gene that codes for GFP (Green Fluorescent Protein). A vector is an agent “employed to transfer the gene from one organism to another” (Lab manual). Two common vectors are phages and plasmids.
Ligation The objective of this experiment was to ligate EGFP DNA inserts into pET41a(+) plasmids. A total of five ligations were performed, two actual ligations and three control ligations. The following reagents were utilized: Nco I/Not I cut pET-41a(+) DNA 50 ng/μL, EGFP cDNA insert 7 ng/μL, uncut pET-41a(+) DNA/EGFP recombinant plasmid DNA 25 ng/μL, ligase buffer 10X, and ligase. To prepare ligation #1, a 1:1 molar ratio of pET41/EGFP, 2 μL digested pET41a(+) DNA, 1 μL EGFP cDNA insert, 14 μL sterile dH2O, 2 μL ligase buffer, and 1 μL DNA ligase were added to a micro centrifuge tube. To prepare ligation #2, a 1:3 molar ratio of pET41/EGFP, 3 μL digested pET41a(+) DNA, 1 μL EGFP cDNA insert, 12 μL sterile dH2O, 2 μL ligase buffer, and 1
However, all proteins are constructed from the same set of 20 amino acids linked in unbranched polymers. The covalent bond that exists between amino acids is called peptide bond, hence a polymer of amino acids is named polypeptide. A protein is a biological functional molecule made up of one or more polypeptides which is folded and coiled into unique three-dimensional structure. In laboratory, it is important to measure the concentration of proteins for research investigations. Biuret test is adopted to quantify proteins in fluid by using a spectrophotometer.
1. Write a sentence for each of these mechanisms describing the manner in which the DNA can be transferred from one cell to another. Transformation: During transformation pieces of genetic instructions are released by a bacterium. Another bacterium, picks up the DNA into its own genome. Bacteria taking up foreign DNA is known as transformation.
Furthermore, Acinetobactor baylyi ADP1 like most organisms undergoes a process known as DNA recombination, where two complementary DNA strands cross and exchange portions of DNA. During recombination, a structure known as a Holliday Junction forms and must be resolved, completing the exchange of DNA (Aravind et al. 2000). Recombination is a crucial mechanism in both gene amplification and deletion. Specifically, ADP1 contains a protein called YqgF, a putative Holliday Junction Resolvase, due to its structural similarity to a known resolvase named RuvC (Aravind et al.
To elute the DNA, the columns were then transferred to 1.5 ml micro-centrifuge tubes and centrifuged at 15 000 g for 2 min. The (quantity and quality of the DNA was tested by spectroscopic measurement (Synergy H4 Hybrid Reader, BioTek) at 260 nm and 280 nm, and the ratio was the criterion for DNA
Epigenetics refers to all modifications to genes other than changes in the DNA sequence itself. This modifications include addition of molecules, like methyl groups, to the DNA backbone. Adding these groups changes the appearance and structure of DNA, altering how a gene can interact with important interpreting molecules in the cell 's nucleus.There are different kinds of epigenetic chemical additions to the genetic sequence. The addition of methyl groups to the DNA backbone is used on some genes to distinguish the gene copy inherited from the father and that inherited from the mother. How do this modifications affect the genes?
The Solid sequencing platform, produced by Technologies/Applied Biosystems (ABI), performs sequencing by ligation method. Similar like the Roche 454 library preparation, genomic double strand DNA were sheared into small pieces and ligated with two types of adatptors P1 and P2 on two ends. One end with P1 adaptor binds onto the surface of the magnetic bead and emulsion PCR takes place to amplify single nucleotide fragment. Then the oil was washed out and four fluorescent labeled di-bases probes were added into the beads mixture. By matching the 1st and 2nd position of the template by di-base probes, fluorescence was detected and the extra tail with fluorescent probe is cleaved out.
What role does the Polymerase Chain Reaction (PCR) play in producing a DNA Profile? PCR amplifies the regions of DNA with short tandem repeats and uses primers with fluorescent labels. This works by replicating the region of DNA several times. The same region is also amplified on both chromosomes, however they are different sizes, which are then put into gel