Plasmid Lab Report

This lab is about moving genes from one thing to another using plasmids. Plasmid has the ability to replicate, so it replicates independently, and separately from the chromosomal DNA. Plasmid are one or more small piece of DNA and they enter cells as a double strand DNA. When they enter the cell as a doubke strand they do not invade he chromosomal DNA. We will also transform bacteria into GFP which is mainly from the jelly fish Aequorea Victoria. The GFP causes the the jelly fish to fluorescent and glow in the dark. After the transformation, bacteria starts to make the GFP which causes them to glow a green color under a ultraviolet light.

The objective of this experiment was to genetically transform E.coli cells to express ampicillin resistance and to produce the green fluorescent protein by using the pGLO plasmid. It was hypothesized that only the cells with pGLO DNA added to the solution would be able to survive in the same environment as the ampicillin, and that only the cells grown in the plate with arabinose would fluoresce bright green. The results supported the hypothesis, showing that the +pGLO LB/amp/ara plate was the only plate that had fluorescent cells and grew. Additionally, that +pGLO cells were the only ones that grew on an LB/amp plate.

This pBlu lab had for purpose to present the changes of the strain of E. coli bacteria due to new genetic information being introduced into the cell. In this experiment we are freezing and heat shocking the E. Coli bacteria that is then forced to take the plasmid DNA. The E. coli then transforms the pBLu plasmid, which carries the genes coding for two identifiable phenotypes. After following the Carolina Biological steps our lab worked well and we able to see some colonies of bacteria on the plates. The x-gal plate showed a significant amount of bacteria to confirm that the pBlu plasmid took over the E. coli strain.

Analysis of DNA from practicals 1 and 2 using the technique of agarose gel electrophoresis and analysis of transfomed E. coli from practical 2 (part B)

The Lambda kit was used to rid the digested pGEM of the stuffer fragment so as to lower the probability of it re-ligating back into the plasmid instead of the luxAB inserts [1]. In total, four ligation reactions were prepared under varying conditions – L1 contained a 1:1 insert to plasmid (with stuffer) ratio, L2 contained a 2:1 insert to plasmid (with stuffer) ratio, L3 contained a 1:1 insert to plasmid (no stuffer) ratio, and L4 contained a 2:1 insert to plasmid (no stuffer) ratio. Strains of highly competent Zippy cells, derived from E. coli DH5 cells, were diluted and transformed with the recombinant plasmids [1]. pGEM contained the Ampr gene which inhibited ampicillin via -lactamase, thus, DH5 cells that took up pGEM were able to actually grow in the agar plates containing ampicillin [1]. In order to determine the efficiency of the ligations and observe any activity of luciferase, several transformations were plated, and the produced blue and white colonies were counted [1]. If pGEM contains an insert, the insert itself will inactivate LacZ by disrupting its sequence -- blue products result from intact pGEM because -galactosidase would properly cleave X-gal, and white products result from cut pGEM because -galactosidase would be non-functional [1]. The negative control was comprised of an aliquot of DH5 cells and sterile water, the positive control was

The possibility of stem cell research exploded with momentum when a certain sheep named Dolly was born. The reason why this was such a breakthrough is because Dolly was the first mammal ever cloned. Although this all began in 1996, the study into stem cell research has been documented since the 1960’s, where Joseph Altman and Gopal Das brought forward new evidence of adult neurogenesis ( the ongoing stem cell activity in the brain). The birth of Dolly brought many breakthroughs for the scientific community but it also created an avalanche of concern because of the ethical implications. With all of the reporting on Dolly, the news media only spoke of one type of cloning and that is reproductive cloning. This may be the most popular

In this lab we learned about plasmolysis. We completed this lab to study the effects of salt and distilled water on elodea cells. We first took a leaf of an elodea plant and placed it on a slide with a drop of distilled water. We then placed a cover slip over the leaf and observed the leaf under a compound microscope with lenses that magnified the cells 100 times and then under a lense that magnified the cells 400 times. We then recorded what we saw by drawing a couple cells in each power. Next, we placed a drop of saltwater on the same leaf and observed it again under the same two magnifications. We recorded what we saw by drawing a the group of cells that we saw. Finally, we added a drop of distilled water to wash out the salt water and repeated the procedure of observing and recording at the two different powers. I knew that salt dissolves in water and that the salt may affect the water levels within the cell. I also knew that when salt was exposed to a plant, the plant wilted. However, I did not have a good understanding of plasmolysis, so I did not have an idea on what was going to happen to the cells during the lab.

Cloning is defined as the “creation of an exact copy of a living matter, such as a cell or organism” according to Encarta encyclopedia. The copies produced through cloning have identical genetic makeup and are known as clones. Scientists use cloning techniques in the laboratory to create copies of cells or organisms with valuable traits.

Plasmids are small double stranded circular non chromosomal DNA molecules containing their own origin of replication. Hence, they are capable of replication independent of the chromosomal DNA in bacteria. Plasmids present in one or more copies per cell, can carry extra chromosomal DNA from one cell to another cell and serve as tools to clone and manipulate genes. Plasmids used exclusively for this purpose are known as vectors. The genes of interest can be inserted into these vector plasmids creating a recombinant plasmid. Recombinant plasmids can play a significant role in gene therapy, DNA vaccination, and drug delivery [Rapley, 2000].

Currently, our society is at a point, where our knowledge and understanding of human, nature, and science are very controversial, yet many promote advancement in scientific technology like cloning as a benefit for humans. Human beings becoming Gods, and manipulating the DNA of animals, plants, bacteria, and humans, so large corporate developers can profit from hybrid creations. Eating from the metaphorical tree of the Knowledge of good and evil, listening to the evil one’s whispers that you shall not die. In addition, people have become smart enough to cause major problems to our society and future as living beings. I believe it is not coincidence that DNA and cloning began advancing around the same time as the Atom Bomb, because both the

DNA is also known Deoxyribonucleic acid. It is the carrier of genetic info. The distinctive characteristics or qualities of someone or something come from DNA. It is regarded as unchangeable. Nearly every cell in a person’s body has the same DNA. A small tiny amount of DNA can also be found in the cell’s mitochondria. Dna is made up of four chemical bases: A,T,C,G. Human DNA consists of 3 million different bases. Each base is also attached to a sugar molecule and a phosphate molecule. Dna can also copy and replicate itself. One strand of DNA is known as the double-helix. It has rungs, just like a ladder. When cells divide, each new cell needs a copy of DNA from the old cell.

In addition to the mph gene, few gene regulations are implemented as safety assessment for the product. In the recombination process of making this product, Ecoli plasmid has been used as a vector to insert mph gene into host bacteria. We use a plasmid vector from E.coli that has conserved sequence for Plac promoter and gef suicide gene. Plac promoter is positioned upstream of mph gene to regulate its expression. Later, Plac promoter will be activated by induction with IPTG.

DNA cloning is the process of creating a multitude of copies of isolated DNA fragments; DNA cloning can be carried out via in vitro or in vivo methods. One can clone a specific DNA sequence or entire gene fragments. There are a multitude of procedures to carry out DNA cloning, but the major steps are the same for all types. To begin the process, one must isolate a DNA fragment from the chromosomal DNA. This is done by using a restriction enzyme. One could also use gel electrophoresis and polymerase chain reaction (PCR). The next step involves introducing the fragment into a DNA sequence that can replicate itself along with the DNA fragment. The restriction enzyme will cut a DNA molecule that can self-replicate, and then the isolated DNA fragment will be inserted by ligation, which connects the fragment to the larger DNA molecule. This new artificially joined DNA is called recombinant DNA.

Imagine a future where humans are manufactured, a future where humans are created by science, a future where humans are the new lab specimen. Human cloning is like opening Pandora's Box, unleashing a torrent of potential evils but at the same time bringing a small seed of hope. No matter how many potential medical and scientific benefits could be made possible by human cloning, it is unethical to clone humans.

Human cloning, long the subject of science fiction, is today a practical reality. Recent breakthroughs, most renowned the cloning of a sheep from an adult cell in Scotland in 1997, have caused the world to acknowledge that human cloning is indeed possible. Governments around the world immediately attempted to address the issue of human cloning, with varying levels of success. At the same time the pace of cloning technology continued to accelerate.