Lab Report On Bacterial Transformation

In this lab, the organism that we have been working with is the bacterium, Serratia marcescens. S. marcescens is a member of the Enterobacteriaceae family, and tends to grow in damp environments. S. marcescens is an ideal bacterium to work with in the lab because it reproduces quicker than other bacterium. This bacterium produces a special pigment called prodigiosin, which is red in color. The prodigiosin pigment is intensified when S. marcescens is grown at higher densities. During our experiment, temperature, pH, salinity concentration and oxygen requirements were tested on S. marcescens to measure their optimal growth and prodigiosin production.

This experiment was designed to test and observe the transformation efficacy of the pUC18 and lux plasmids in making E. coli resistant to ampicillin. Both plasmids code for ampicillin resistance, however, the lux plasmid codes for a bioluminescence gene that is expressed if properly introduced into the bacteria’s genome. The E. coli cultures were mixed with a calcium chloride solution and then heat shocked, allowing the plasmids to enter the bacteria and assimilate into the bacterial DNA. The plasmids and the bacteria were then mixed in different test tubes and then evenly spread onto petri dishes using a bacterial spreader, heating the spreader between each sample to make sure there is no cross contamination. Each of the dishes was labeled and then incubated for a period of 24 hours. The results were rather odd because every single one of the samples grew. Several errors could have occurred here, cross contamination or possibly an error in preparation as every single sample in the class grew, meaning all samples of the bacteria transformed and became ampicillin resistant.

The purpose of this experiment is to make E.Coli competent so that it can be transformed in order to become immune to ampicillin, then we would be able to determine the transformation efficiency of the culture. We determine this by preparing 4 plates of E.coli, each labeled “LB-plasmid”, “LB+plasmid”, “LB?Amp-plasmid”, and “LB/Amp+plasmid”. This meant that either should have lacked plasmid and Ampicillin, with plasmid but lacked Ampicillin, without plasmid but with Ampicillin, or were with Ampicillin and plasmid, respectively. Then we made the bacterial cells competent by adding CaCl2 to 2 vials of the colony (one with plasmids), and incubating on ice, then heat shocking, and returning to ice. Luria Broth is then added and left to sit for 5-15

The purpose of this experiment was to show the genetic transformation of E. coli bacteria with a plasmid that codes for Green Fluorescent Protein (GFP) and contains a gene regulatory system that confers ampicillin resistance. A plasmid is a genetic structure in a cell that can replicate independently of chromosomes. In this lab, the Green Fluorescent Protein, which is typically found in the bioluminescent jellyfish Aequorea Victoria, was cloned, purified, and moved from one organism to another with the use of pGlo plasmids. It was hypothesized that if bacteria that were transformed with +pGlo plasmids are given the gene for GFP, then transformed cell colonies

One of the most imperative functions in maintaining the development of evolution is the frequency of genetic transformation: the injection of foreign DNA into another organism’s DNA. This term is defined by the actions of a vector, but more specifically by the actions of plasmids and phages. However, in this experiment we are primarily focused on the effect of the pGLO plasmid transformation of GFP on the E. coli bacteria by introducing a second chromosome or a plethora of cloned plasmids. (Bassiri 2011)

For this experiment, E. coli was best for genetic engineering because of their size, and their fast reproduction (Spilios, 2017). E. coli will be genetically transformed using an engineered plasmid. A plasmid is a circular piece of DNA which independently replicates and multiplies because it has its own origin of replication (Spilios, 2017). The pGLO is the plasmid used in this experiment. Plasmids are used as vectors and they contain manipulated genes such as genes coding for antibiotic resistance for drugs like ampicillin. This antibiotic resistance of such serves as the selectable marker in genetic transformation and for genetic transformation to proceed, the cell must reach competency which is the physiological state that is required for the vector plasmid to get into the cell for transformation (Spilios, 2017). While competency can be reached naturally in some organism, it must be reached artificially in E. coli through treatment with CaCl2 and exposing them to heat shock using incubation (Spilios, 2017).

If a gene that codes for Green Fluorescent Protein transforms bacteria and GFP glows when transformation occurs, then when two micro test tubes have 250 microliters of transformation solution and places in an ice bath, then 2-4 bacteria colonies are added to each tube with a sterile loop; then a plasmid (pGLO) is added to one of the tubes, incubated in ice for 10 minutes, then heat shocked for 50 seconds at 42 degrees Celsius, then back into 9ice for two minutes; then LB nutrient broth is added to both tubes (250 microliters) and set out at room temperature for 10 minutes. Then, 100 microliters of each solution in the tube are added to four

After analyzing the data recorded for both the agar plates containing ampicillin and those that did not, it can be concluded that the data provides enough evidence to reject the null hypothesis. There is enough evidence to support the alternative hypothesis stating that there is a correlation between plasmids coding for an antibiotic resistant gene and bacterial growth in ampicillin. When a bacterial solution containing either pUC18 or the lux plasmid is transformed in an agar plate containing ampicillin, only those cells which took in the plasmid are able to survive and replicate, forming individual colonies. Not all cells are transformed though, the chances of a successful transformation were extremely low. In order to see which cell transformed the cells were tagged according to their plasmid, in the plates containing pUC18 the only

The purpose of this experiment is to show how different concentrations of Ampicillin affect Escherichia Coli Growth and how the bacteria become resistant to the antibiotic drug. Through a series of steps, which involves streaking agar plates with E.coli sample and application of ampicillin to the E.coli sample on the agar plate, the experiment yields a result that supports the hypothesis. The hypothesis acclaims that ampicillin would affect the growth of E.coli; measuring the zone of inhibition approves the claim in the experiment. The measurements of the zone of inhibition indicate that the generation of E.coli expands as the radius reduces. The reduction of the radius shows the E.coli population is reducing and becoming resistant. In other

Transformation is a procedure where cells take up foreign DNA from their environment. This can happen in either prokaryotic or eukaryotic cells. A circular self-replicating molecule of DNA known as a plasmid transfers genetic information between bacteria enabling the microbes to gain antibiotic resistance as well as the ability to adapt to a new environment. Bacteria also have the capability to acquire small pieces (at random) of DNA from their environment but this however is said to be inefficient because only a small fraction of cells will uptake DNA and become transformed. Expressions of genes on the plasmid will alter the genotype and phenotype of the cell. Under specific laboratory conditions a cell which is incubated with the plasmid DNA has the ability to absorb the plasmid into its own cytoplasm. Because these natural processes can potentially be modified by scientists, the transformation procedure can be used as a means of treating human diseases to increase the quality of life.

In genetic transformation, vectors are used to transfer genes from organism to another. In this experiment, we used plasmids as

If genetic transformation allows the insertion of a gene into an organism to gain the trait that the inserted gene codes for, then when 250 µl of transformation solution(CaCl2) and 2-4 colonies of bacteria, E. coli, are added to two micro test tubes, labelled +pGLO and –pGLO, with a loopful of pGLO DNA solution in the +pGLO tube, and are incubated for 10 minutes, then transferred to a 42ºC heat bath for exactly 50 seconds, and then incubated in ice again for 2 minutes for a heat shock, and then have 250 µl of LB nutrient broth, and lastly 100 µl of the solutions spread across the surfaces of 4 LB nutrient agar plates, LB/amp, LB/amp/ara, LB/amp, and LB, with +pGLO, +pGLO, –pGLO, and –pGLO used, respectively, the plates with the –pGLO solution will not glow under a UV light after they have

This report discusses an experiment which students have to transform and plate competent Escherichia coli in different concentrations of plasmid DNA. This experiment uses four concentrations of plasmid DNA to perform four transformations. These concentrations are namely, 5 µl of distilled water, which acts as the control in this experiment; 2.5 µl of undiluted plasmid DNA; 1.0µl of undiluted plasmid DNA; and lastly, 1.0µl of plasmid DNA diluted 1 in 10. Transformation is an important aspect in genetic engineering as it allows for a particular DNA to enter into another cell. The aim of this experiment is to be able to perform a cell transformation

In genetic transformation, DNA genes of one organism are added to another organism. The DNA that has been transmitted to the new recipient of the genetic material is then expressed and continued (Holmes et al. 1996). By doing this, large quantities of preferable genes can be made. The process of transformation can be done naturally or artificially. Escherichia coli is a common bacterium this is single celled allowing the process for plasmid exchange to not be difficult. The bacterium produces clones allowing the DNA to not be mixed.

Bacterial transformation plays a huge role in this lab. Bacterial transformation is the process by which bacterial cells take up naked DNA molecules. If the foreign DNA has an origin of replication recognized by the host cell DNA polymerase, the bacteria will replicate the foreign DNA along with their own DNA. The field of study that uses bacterial transformation as their driving force for research is biotechnology. The lab that was carried out in class is just a very small example of biotechnology. Biotechnology according to www.bio.org is just simply technology based on biology that harnesses cellular and bimolecular processes to develop technologies and products that help improve our lives and the health of our planet. This field of study is used all over the world to provide for solutions for many problems that previously were not able to be solved. Some examples of areas where this technology might be used, range from the medical field, all the way to the other side of the spectrum in the agriculture business. Many patients are given the opportunity to be treated with different kinds of medicines that would not be possible without biotechnology. An example of this would be human insulin. Some people are not able to make their own insulin, this disease is called diabetes. The human insulin gene is injected into these bacteria and as a result these bacteria make human insulin which is then harvested and sold for the people that can’t make