The Community of Microorganisms that Reside in the Epithelia of Humans

A healthy adult has over 1000 species of bacteria residing in the gut (1). There is great diversity in genera of these bacteria, however the most

Often scientists work with bacteria that do not come in a labeled test tube— for example, bacterial samples taken from infected human tissue or from the soil—and the scientist must then identify the unknown microorganism in order to understand what behavior to expect from the organism, for example, a certain type of infection or antibiotic resistance. However, because of the relatively few forms of bacteria compared to animals and because of the lack of bacterial fossil records due to their asexually reproductive nature, the taxonomy used to classify animals cannot be applied to bacteria (Brown 275). In order to classify unknown bacteria, a variety of physiological and metabolic tests are available to narrow a sample down from the fathomless number of possibilities into a more manageable range. Once these tests have been performed, the researcher can consult Bergey’s Manual of Determinative Bacteriology, a systematically arranged and continually updated collection of all known bacteria based on their structure, metabolism, and other attributes.

Bacteria are ubiquitous; they can be found on the skin, in the soil, and inside the body. Because of the very nature of this ubiquity, it is important to be able to determine between different strains of bacteria. An example of this is determining the causative agent for a disease so that the patient will be treated with the appropriate antibiotics. It may be important to determine the bacteria in a certain region, because like with enteric bacteria, it is normal to find them in the digestive tract as they are in a symbiotic relationship with our bodies in this area; however, they also cause opportunistic infections in places outside of the digestive tract to our detriment, such as with a urinary tract infection. Some strains of bacteria are common to nosocomial infections, and identifying these bacteria as such helps create the guidelines for healthcare workers in antiseptic technique. All of the morphology and characteristics of each strain of bacteria help us to better understand the role of bacteria in the body as well as helps us understand how they can cause illness, and what treatment regimen to set in place. In lab this semester, a sample of unknown

PCR is the amplification of DNA by denaturing, annealing, and extension of a DNA template. Specific sequences can be amplified using single-stranded DNA that complements the target sequences known as primers. This process heats DNA until the strands separate, then primers bind to the target regions. DNA polymerase enzymes and single base nucleotides (dNTPs) are used to synthesize new strands of DNA to the target sequence. The end product will contain large quantities of the target sequence (Bean et al. 2015). The most notable in phylogenic studies is the 16S rRNA gene, because of it’s highly conserved primer-binding site and hyper variable regions that provide species-specific sequences within bacteria and archaea (Kolbert and Persin 1999). This gene is a component of the 30S small subunit of prokaryotic ribosome’s and serves as the primary site of protein synthesis. (Woese and Fox 1977). The 16S rRNA sequence can be amplified and matched to national databases provided by the National Center for Biotechnology Information (NCBI) using software termed Basic Local Alignment Search Tool (BLAST) to find regions of similarity between biological sequences for bacterial identification. Thus, providing a cost effective and timely method when compared to biochemical

Different microbes can transmit and produce different types of diseases and infections. Having an unknown bacterium in the body can be a life and death situation. It is very important especially in the healthcare industry that providers are able to differentiate between organisms that are pathogenic and administer the appropriate treatment to their patients. Applying methods that were previously studied in lab, students must be able to isolate an unknown specimen by using laboratory techniques and biochemical tests.

Unknown lab report# 24 Introduction Microbiology is a study of organisms that surrounds us every day. It requires an educational understanding to identify organisms, and the ability to distinguish different bacteria’s. In applying the learning process of the different bacteria’s, unknown bacteria samples are given to be studied and identified.

The struggle of finding accurate technology to be able to compare human microbial communities globally include the degree of relatedness so whether that persons are related, the place in the family structure, the age, demographics, there are many ethical, legal, and logistical barriers, types of comparisons,

A highly conserved gene will be used to identify a prokaryotic species isolated from the body. Fundamental lab techniques will be also explored and utilized, such as amplifying using PCR, cloning, and transforming the gene into a host cell. DNA electrophoresis and specific substrate plating will serve as analysis check points. The final product will be sequenced and compared to similar species to observe phylogenetic relationships.

The purpose of the experiment was to isolate plasmid DNA, followed by restriction digestion using restriction endonucleases and then visualizing the digested fragments after subjecting to gel electrophoresis. Plasmid DNA (pSP72 DNA) was isolated from Escherichia coli KAM32 (E.coli) cultures using the QIA prep miniprep kit and then subjected to restriction digestion by EcoRI and HindIII. The restriction digested DNA was then loaded into the wells of 0.7% agarose gel and subjected to electrophoresis. It can be concluded from our results that our plasmid DNA isolation was successful and the restriction digestion results were partially in agreement with our hypothesis.

In this laboratory experiment, the students underwent a series of different lab techniques starting off with the process of PCR reaction and electrophoresis by examining their cheek cells and using the cells to show the Alu repeats between the DNA of their laboratory groups. The students began by obtaining cheek cells with toothpicks, putting them in a thermocycler, creating a PCR reaction with these cells, and finally using the amplified DNA to understand the repetition of Alu in the gel substance in electrophoresis. The portion of the laboratory experiment that was non-controllable by the students was the evaporation of some group member’s PCR reactions. The micro centrifuge tubes sat for a week until the next lab period, some of the PCR

Isolation of plasmid DNA from three cultures of E.coli using a method known as the alkaline lysis method.

After the incubation, 1.5 mL of each of the three cultures were added to eppendorf tubes and centrifuged at 13,200 rpm for 1 minute. An alkaline lysis procedure like that of Birnboim and Doly was then performed to extract the plasmid DNA with 200 μl of alkaline SDS detergent solution (Birnboim & Doly, 1979). After

Researchers studying microorganisms’ community use genomics recognize myriad factors and interactions help shape the composition of the microorganism. However, in different people, the microbe varies in development at various times with relation to the disease. Infectious research on the diseases is dominated continuously by reducing the approaches (Casadevall and Pirofski). The microbiologists view the microbes of any disease as the major factors that vary with the disease. They, therefore, treat the body of the host as just a

The chemical and reagents used for the extraction and quantitation of DNA were: Plant DNAzol (0.3ml/0.1g), 100% ethanol (100%: 0.225 ml/0.1 g, 75%: 0.3 ml/0.1 g), Chloroform (0.3 ml/0.1 g), Plant DNAzol-ethanol solution: Plant DNAzol, 100% ethanol (1:0.75 v/v), TE buffer (10 mM Tris, 1 mM EDTA pH 8.0), 1.2% agarose gel (Agarose, 1X TAE buffer), 6X loading buffer (glycerol, Tris/EDTA pH 8.0, ethidium bromide), .25X TAE buffer, Restriction enzymes and Restriction endonuclease buffers. All the chemicals used were quality grade. The restriction

This essay based on the principle of real time PCR which uses CYBR green dye that combines to any double stand DNA. This process included two maim steps. The first step was designing of HSV-1 primer and /or HSV-2 primer (we have chosen only HSV-1). BioEdit software has been used to edit the nuclide sequences where necessary. After that the primer has been ordered. The second step was in the laboratory which included applying the HSV-1 primers to real time PCR protocol. The final results of this protocols aim to demonstrate the quality of HSV-1 primer that we have designed by interpretation three criteria. These are specificity, efficiency and