Flow Diagram Lab Report

The next step of the project included preparing a Gram stain to discover the cell shape, arrangement, and if the bacteria is gram positive or

There are many reasons for knowing the identity of microorganisms. The reasons range from knowing the causative agent of a disease in a patient, so as to know how it can be treated, to knowing the correct microorganism to be used for making certain foods or antibiotics. This study was done by applying all of the methods that I have been learned so far in the microbiology laboratory class for the identification of an unknown bacterium.

2) Record the shape of the bacteria, the arrangement of the bacteria, and the gram staining characteristics.

Many tests were completed on the unknown such as gram staining and inspection under microscopes to find whether the bacterium is gram positive or gram negative. Chemical resistance tests were also performed to see if certain chemicals affected the unknown growth or if it didn’t affect the bacteria at all. Each biochemical test

134). They are loops of DNA that are separate from the chromosomal DNA and can self-replicate in a cell, found mostly in bacteria (Brown, 2011; Addgene, 2015). Lederberg and William Hayes discovered that plasmids were being transferred from one cell to another, not the chromosomal DNA (Brown, 2011, p. 135). This discovery lead to plasmids being an essential tool for scientists. Scientists can engineer plasmids to have specific genes to introduce into new cells (Brown, 2011, p. 134). On a plasmid loop there will be an origin of replication (ORI) and a multiple cloning site (MCS) where the gene of interest is inserted (Bio-Rad, 2015). This region has specific restriction enzyme recognition sites, which are cut by the enzymes to open up the DNA where the new gene will be inserted (Jove Science Education Database, 2015). Most plasmids will also contain an antibiotic resistance gene allowing cell survival in environments containing antibiotics (Jove Science Education Database, 2015).

The identification of unknown organisms carries important ramifications that can be applied to many real world scenarios. In keeping with quality assurance beverages, food, cosmetics, and other products are frequently inspected for contaminants resulting from a presence of pathogenic bacteria. In medicine, a physician’s diagnosis and consequent treatment is largely determined from samples collected from infection sites that have been analyzed using microbial tests.

Lab Day 1: After receiving my unknown bacteria, I streaked a TSA plate and incubated at 37°C for 48 hours. I then picked a single colony from the plate with my sterile loop and streaked a TSA slant and labeled it “Working Stock”. I did the same with another TSA slant and label the second one “Back-up Stock”. This would be the samples I used to complete the following procedures through the next four weeks to determine my unknown bacteria.

is done to confirm plasmid identities by their amount and size and to examine unknown plas-

The objective of this experiment is to identify the organisms of two unknown bacterial cultures. Students must identify the species of the unknown bacteria by utilizing the techniques and information learned in previous laboratory exercises. These techniques include streaking for isolation, Gram staining, and specific biochemical tests. Students are given a map known as a dichotomous key, a guide in determining the identity of their unknown sample.

Our research on recombinant DNA mainly consisted of two experiments: Transformation and gel electrophoresis. In our first experiment, four microfuge tubes were given to us: pKAN DNA, pAMP DNA, unknown DNA, and a TE buffer without DNA. The two positive controls, pKAN and pAMP, consisted of an antibiotic resistance gene respectively to their name. The pKAN plasmid contained the gene resistance for kanamycin while pAMP carried the gene resistance for ampicillin. The negative control, TE, only contained buffer without DNA. The fourth tube was our unknown plasmid, which was either pKAN or pAMP; and by way of artificial transformation, we would be able to initiate the identification of our unknown plasmid.

: In the field of microbiology, there are times when a sample will contain more than one species of bacteria. The goal is to separate each bacterium and culture them independently from one another to identify them. This was the objective of this lab. Each stock contained two unknown bacterium, and the possible unknowns were Eserichia coli, Enterobacter aerogenes, Enterobacter cloacae, Serratia marcescens, Proteus vulgaris, Klebsiella pneumoniae, Shigella flexneri, Shigella sonnei, and Salmonella enterica. The tests available were MacConkey agar, Endo agar, Hektoen Enteric agar, Tryptone Soya Agar (TSA), carbohydrate sugar broths, Triple Sugar Iron (TSI) agar, decarboxylase broths (arginine, lysine, and ornithine), Simmon’s Citrate Agar, urease

Previously, detection and recognisation of bacteria were based on conservative tube-based biochemical reactions, and their results were compared to historical charts of expected biochemical reactions. Beacause of the need for quicker, simpler methods,

During the course of this lab, we have learned to utilize the flow process chart to break down the operation into its basic elements. We then made observations of the chart, in order to determine recommendations for increasing the efficiency of the operation. In the course of our operation we were able to determine the average weight of the three cell phones to be 201 g.

Another purpose of this experiment is to stress the importance of knowing the identity of a microorganism. Knowing the species of microorganism present in a sample provides a

In the second article Mcgarvey, Queitsch, and Fields are attempting to study and identify which genes and or proteins are accountable for generating antibiotic resistance in bacteria. The authors hypothesized that a screening of soil metagenomic DNA library in an Escherichia coli host for genes could project resistance to the following antibiotics: gentamicin, kanamycin, rifampin, trimethoprim, chloramphenicol, or tetracycline. The screen concluded that 41 genes were encoding novel protein variants of about eight families.