No quick endospore stain was performed to validate this assumption since only one assigned organism was endospore forming and unlike Unknown #10, that organism was Gram positive. By Gram staining alone, it was safe to eliminate the three Gram positive bacteria that could have been assigned: S. epidermidis, M. luteus, and B. megaterium. The second step was to streak plate Unknown #10 to observe its macroscopic
GFP was needed so that we would tell if the ampicillin resistance gene had been properly placed when the bacteria glowed under a UV light. The purpose of this lab was to perform a procedure known as genetic transformation which allowed us to genetically engineer E. Coli to be ampicillin resistance. Before the lab we expected that lysogeny broth and minus DNA will have growth but no glow. The lysogeny broth, ampicillin, and
After receiving an unknown mixture, the sample was streaked for isolation onto TSA, blood agar, and MacConkey plates. Each plate serves as a first step to identify the unknowns. The TSA (tryptic soy agar) can be used to do a gram stain, which differentiates gram-negatives from gram-positives, based on the structural make up of the cell wall (Carson, 2015). The blood agar plate is used to test for hemolytic activity, which is useful for distinguishing gram-positives. A MacConkey plate is selective by inhibiting the growth of gram-positives and differential due to the fermentation of lactose by certain gram-negative species.
Chromosome #2 with locus TPOX with a length of 248 base pairs was only found in the parent sample A. None of the chromosomes from sample b matched the chromosomes from A, C, or D+E except for maternal TPOX chromosome from B and the paternal TPOX from C. 3. Would this exercise still work properly if you had chosen any combination of maternal and paternal chromosomes for chromosomes 2, 5, 7, and 13 from samples D & E? Yes because the maternal and parental chromosomes would still be put on the electrophoresis table. The parents turn out different based on DNA combos that are chosen.
According to the series of test that my group ran for our unknown specimen, we had a match with the bacteria known as Alcaligenes Faecalis. This bacterium belongs to one of the major group of gram-negative bacteria (Phylum Proteobacteria). Alcaligenes Faecalis (Genus, species) is a rod shaped (bacillus), 0.5-1.2 x 1.0-3.0 µm, round with scalloped margin (colony configuration growth), motile (with one to nine peritrichous flagella), gram-negative, non-fermentative bacteria, obligate aerobic, having oxygen as the principal terminal electron acceptor in the electron transport chain (ETC). We consider we have a match with the species Alcaligenes Faecalis because of the following reasons: Fermentation tests performed (Durham sugars) were negative, which indicate that our bacteria use a different metabolic means for growth (non-fermentative gram-negative bacteria).
This is due to the fact that n order for the E coli. bacteria to gain either of the traits the plasmid had to be present in the first place, because the GFP gene is inside the plasmid. If arabinose is not in the media in which the bacteria was growing on then the GFP gene could not turn on, thus the bacteria can not glow. This is why the LB/amp/ara plate was the only one to express both traits(antibiotic resistance and glowing). The second part to our hypothesis was that using HIC,GFP would be purified and the final tube would express GFP under an ultra violet light.
The unknown bacteria was then tested on multiple selective and differential media. Growth was present on the MacConkey Agar and the colonies were the same color as the plate, which told me my bacteria was gram negative and did not ferment lactose. There was no growth on the Mannitol Salt Agar, and this told me the unknown was not salt tolerant and did not
Our results from the PCR process were very unexpected, even to the point the control colony had some rather odd outcomes. The goal of this experiment was to choose three colonies from the petri dish that has been exposed to +Amp, and look for any signs of the +Amp resistant gene, blaTEM, within the colonies and decide if this gene does have an impact on bacterial resistance towards the antibiotic. My partner and I decided to utilize a bacterial colony sample that does have blaTEM genes as our control group for us to indicate what a blaTEM gel strand would appear in the agarose gel results. When observing the product of the gel product after gel electrophoresis, we were surprised to find out none of our three colonies had any strands that indicated the presence of blaTEM despite each of them surviving through the exposure to this antibiotic.
although, we had tested Ag+ we had no precipitate form in our solution. accordingly, we did not have to test for Ag+ since no precipitate formed and skipped the entire process for Ag+. carrying on to the next step we had to test for iron. Before we can start testing for iron. I had to prepare the solution, preparing the solution contained taking the supernate that we made from group one and adding the following; NH4Cl, NH3, water and (NH4)S to the solution along with adding
Post Lab Questions for Immunofluorescence 1.) In our experiment the primary antibody that was used was anti-tubulin, which was generated in the rabbit anti-mouse tubulin antibody. It is specific for the tubulin proteins to help identify their binding sites. 2.) The secondary antibody that was used was anti-actin, which was stained with FITC.
The repressor is a regulatory protein that binds to the operator and blocks transcription of the genes of an operon. Inducers bind to the repressors and they also regulate gene expression. In the process of identifying the three strains of E.coli, ONPG (ortho-nitrophenyl b-D galactoside) was used as an indicator. ONPG is a substrate that can detect B-galactosidase, and when it does, it turns yellow. Sarkosyl was also a detergent used in the lab to lyse open cells.