PGLO Write Up Purpose The full on purpose and meaning of the PGLO lab was to see and watch transformation happen and to understand it. Transformation is when there’s a genetic alteration in a bacterial cell, or a alternation in a cell. To get into full detail of what the purpose was it was to Transform the E-coli bacteria as used in the lab to glow green by adding plasmids, then it should be enlarged in bacteria colones. When lighten up by a UV light the bacteria should glow green. Procedure The PGLO lab itself was fairly easy, but don’t get me wrong there were a couple of steps you had to be very precise on. The first few steps are fairly easy you had to label your test tubes then filling two of the +PGLO tubes with your transformation …show more content…
#2 For agriculture transformation is used for coding certain traits. #3 it can gene transfer or that's what it can do. #4 It can help make insulins, or growth hormones, and its used to genetically change bacteria. #5 It is because plasmids move cell to cell extremely easily. #6 It’s when the bacteria evolves to the drug and then becomes useless. #7 The plasmid is ampicillin. #8 it is arabinose. #9 a single cell organism, because it can reproduce itself and give the clone of itself the same genetics. #10 slowly because you can see the traits more slowly and get to see the traits change. #11 It should be a harmless organism in the lab. #12 Bacteria because it’s a fast growing and changing organism. #13 you can grow it on the agar plate and watch it grow. #14 it stops the e.coli cells from killing bacteria. #16 on the +DNA plates because dna the plasmid. #17+DNA plates because the plasmids. What I expected to find from my knowledge from transformation that it would grow bacteria on the plates and it would glow. Data For the +PGLO LB/AMP there was a great amount of growth on the culture plate. For the +PGLO LB/AMP/ARA there was as significant amount of growth and it Glowed. The -PGLO LB had an extreme amount of growth on the plate, and for the -PGLO LB/AMP there was no growth …show more content…
#21 They were different, because the -DNA LB plate doesn't have the plasmid. #22 It is that only the one with the +DNA/LB/AMP/ARA was the one that the GFP gene switches causing the bacteria to glow green. #23 one had DNA and one doesn't have DNA. #24 It’s the bacteria growing. Analysis part 2 Questions #1 One the traits that was not altered in all was the color. #2 We proved it by shining a UV light or Ultraviolet light onto the culture plates to see which one glowed. #3 The GFP gene was the gene that made the glow in the bacterial colonies. #4 The evidence we have is seeing if it glows or not, if it does glow you were successful if not you failed. #5 The PGLO plasmid used as a vector is what makes it glow with the fluorescent protein it carries. #6 The organism itself can save energy by being able to turn genes on and off. Conclusion Know in the modern time bacterial transformation is used in many ways such as, making medicines, insulins, cloning DNA and excetera. It is very useful and is used all the time everywhere in the world. Transformation itself is a very useful method, but can also be very harmful if used to making something bad. Which raises big red flags for some people and can be a ethical problem indeed. Using bacterial transformation is good though. It is used in dozens of ways for the good and scientific studies and we should use it to our advantage
Digestion of the haemolytic and non-haemolytic cells allowed for easier identification of fragments during electrophoresis analysis. Lane 12 in figure 3 show the size markers of SPP1 digested with EcoR1 while lanes 6 and 7 show samples of pK184hlyA and pBluescript digested with EcoR1 and Pst1. Lane 4 was loaded with plasmid DNA from haemolytic cells digested with EcoR1 and Pst1 while lane 5 was loaded with EcoR1 and Pst1 digested DNA from non-haemolytic cells. There was a lack of technical success in both lanes due to no bands appearing in lane 4 and only a single band appearing in lane 5. Theoretically, two bands should appear in both lanes after successful to allow for fragment identification. A possible explanation for the single, large fragment in lane 5 is that successful digestion did not take place and the plasmid was only cut at one restriction site leaving a large linear fragment of plasmid DNA. The absence of bands in lane 4 could be because there was not enough plasmid loaded into the lane. Another possibility could be that low plasmid yield as obtained when eluting the experimental samples in order to purify it. Lanes 8 and 9 belonged to another group and show technical success as two bands were present in both the haemolytic (lane 8) and non-haemolytic (lane 9) lanes. If the
Other human errors could have affected the results, such as not inverting the plate before putting it into incubation would not allow for bacterial growth. Not pipetting the tube up and down to mix the bacteria that settled at the bottom of the tube before starting the Gram Stain would not allow for an accurate reading because there wouldn’t be many bacteria on the slide. Passing the slide over the bunsen burner too many times, hence killing the bacteria and not allowing for a Gram Stain. If this experiment had to be redone, one improvement would be to allow for more than one plate without a point deduction. Unexpected human errors might interfere with a person’s results.
Therefore colonies containing the non-recombinant pUC19 plasmid have a functional lacz’ gene appear blue on the agar and colonies containing recombinant pUC19 would have a non-functional lacz’ gene due to insertional inactivation and appear white on the growing medium.
The main goal for our experiment was to learn how to examine DNA when there is only a small
It has an outer membrane that contains lipopolysaccharides, a periplasmic space with a peptidoglycan layer, and an inner cytoplasmic membrane. It also consists of adhesive fimbriae. Some strains of E. coli are piliated and are capable of accepting, as well as transferring plasmid to and from other bacteria. This enables the bacteria under stressful or bad conditions to survive. Although its structure is simple with only one chromosomal DNA and a plasmid, it can perform complicated metabolism to help maintain its cell division and cell growth. E. coli produce very rapidly; a single microscopic cell can divide to form a visible colony with millions of cells overnight (phschool.com). It is the preferred bacteria in most laboratories because it grows fast and easy, and can obtain energy from a wide variety of sources. Since the birth of molecular cloning, E. coli has been used as a host for introduced DNA sequences (biotechlearn.org.nz). In 1973, Boyer and Cohen showed that two short pieces of DNA could be cut and pasted together, and returned to
Hypothesis: If a GFP gene is inserted into an E.coli cell, then the E.coli will glow in the dark.
I described and applied physiological and biomechanical concepts related to physical activity and skill in EXS 397 lab. A student in the lab was tested on their VO2 max using The Bruce Treadmill protocol. With the data I recorded from the test, I was able to apply physiological and biomechanical concepts to explain the subject’s energy sources during the run.
Coli can in fact undergo mitosis on plates containing ampicillin, and show fluorescent qualities on a plate containing arabinose. These results logically follows from the fact that the plasmid inserted possesses qualities that allow for ampicillin to be broken down, and therefore not harm the E. Coli, meaning growth on plates containing ampicillin is proof of genetic transformation. Similarly, transformation of the E. Coli is also evident on the plate containing arabinose, as there was not only growth, but clear fluorescence under the blacklight, as the plasmid also codes for that expression. It is clear that those results are a result of the plasmid, as the plates treated with the +pGLO solution can be compared to those with the -pGLO solution, in which there was no growth on any plate except for the LB broth plate. Growth on the LB plate indicates that the E. Coli is healthy, and capable of mitosis in certain conditions, but lack of growth on the other plates points to it still being wild type. Therefore, it is clear that the +pGLO E. Coli have adopted new genes that allow for new functions that wild type E. Coli are incapable of, in addition to showing that the genes were transcripted and translated in S phase of mitosis, as daughter cells possess similar qualities, as shown by their ability to subsequently grow and divide. This In further examining the plates treated with the +pGLO solution,
The scientific techniques that were mainly used in the conduction of our investigation were the identification of bacteria through the use of a microscope, putting together a Simple Distillation Kit, treating Petri Dishes with Agar on them with the boiled, distilled or radiated water and calculating the coverage of bacteria on the amount of area treated on the Petri Dish. All of these scientific techniques were a crucial part of our investigation. Without all of these techniques we wouldn’t have been able to conduct such a successful experiment.
Transfer 100 µL of cells from the “-plasmid” tube to the agar plates marked with “LB/Amp -plasmid” and “LB -plasmid.” Take an inoculating loop and bend one of the ends into a 90 degree angle so that the loop now resembles a hockey stick. Use the bent inoculating loop to spread the cells along the plate’s surface while making sure that you are not piercing the agar’s surface. Repeat the same process when transferring cells from the “+plasmid” tube to the plates labeled “LB/Amp +plasmid” and “LB +plasmid.”
The purpose of Scientific Method Lab is to determine which variable mass or height will have the greatest impact on the speed of a sphere. The generalized purpose of this lab is to practice scientific method and to learn how to write lab procedures.
Green Fluorescent Protein fluoresces bright green when exposed to UV light. The GFP gene only activates if there is arabinose present. When arabinose is not present, the arabinose digestion genes are inactive and energy will be conserved. However, when arabinose is present the genes activate and start to break down the arabinose until it is all consumed. BAD encodes for the enzymes used to digest arabinose. The araC gene in the DNA map above pairs with arabinose, and up regulates BAD. However, it also negatively feeds back into its own regulation. Bla is the Ampicillin Resistance gene produces Beta lactamase, a protein that confers ampicillin resistance [1].
I predict, that after 24 hours, that the E. Coli that took up the pGlO plasmid will have the GFP gene and with the GFP gene if there is sugar present I believe that you will see the bacteria glow in the dark. Only the cells that take up the pGLO plasmid will survive when ampicillin is present because the ampicillin will kill the bacteria without the pGLO. The results of this experiment are important to know because, you can find out if you had an error in your procedure. Also if you know the results you can sometimes maybe you can leave out some of the experiments you are trying because you know they will not work. We know the results from what we read in the lab
Genetically modified organisms have also been developed for commercial use. Perhaps the most famous example is the food crops, such as soybeans and corn, genetically modified crops and genetically modified crops. Scientists have studied “knock-out” mice，which are transgenic mice that have a particular gene of interest disabled. By studying the effects of missing genes, scientists can better understand the normal function of the gene and how can we improve the uses of transgenic technology. The more we know about genes, scientists can create more useful things for human beings through transgenic technology.
The whole idea here is to insert the gene and find out the one that work and separate them from the null ones, then reproduce them in a large amount and the final stage will be the verification of what we want. The last step might be overlooked by many people, but oftentimes is more important than we think it will be. Because even if the desired traits have been inherited through the selection, but they might perform differently under different environmental conditions and.