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
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#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
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#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
The unknown bacterium that was handed out by the professor labeled “E19” was an irregular and raised shaped bacteria with a smooth texture and it had a white creamy color. The slant growth pattern was filiform and there was a turbid growth in the broth. After all the tests were complete and the results were compared the unknown bacterium was defined as Shigella sonnei. The results that narrowed it down the most were the gram stain, the lactose fermentation test, the citrate utilization test and the indole test. The results for each of the tests performed are listed in Table 1.1 below.
The first step of the experiment was ligation, and the objective was to insert EGFP cDNA into a restriction cut pET41a(+) vector to obtain a recombinant plasmid that would express green fluorescent gene. pET41a(+) was the choice of vector to ligate the EGFP into. Its structural design and genomic sequential properties render it especially well-suited for cloning and high-level expression of peptide sequences. This 5933 bp circular vector contains a built in sequence for Kanamayacin resistance gene. “Rooting of non-transgenic shoots was completely inhibited in all culture media containing kanamycin” (Montserrat, et. al., 2001). This allowed the growth of recombinant and non-recombinant colonies of E. coli, all of which contained the vector insert.
The plasmids in lanes 3,4,8 and 9 have been digested using one restriction enzyme and had been cut at one restriction site, resulting in a linear molecule. Comparing lanes 3 and 4 to
The purpose of this experiment is to identify an unknown insert DNA by using plasmid DNA as a vector to duplicate the unknown insert DNA. The bacteria will then be transformed by having it take in the plasmid DNA, which will allow us to identify our unknown insert as either the cat gene or the kan gene.
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 gel electrophorese, 2.5ul of the 10x loading dye is added to each PCR reaction tube. The gel
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
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.
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.
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,
loop. Spin the loop in the tubes’ solutions to make sure that all of E. coli’s cell mass is
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.