What Is A Transposon?

As predicted the E. coli colony transformed with either the PUC18 or the lux plasmid developed an ampicillin resistance. Which made it easier for them to not only survive but also replicate in both the LB agar plates and the LB ampicillin rich agar plate. However the E. coli colony not treated with the plasmids could not survive and colonize in the LB ampicillin rich agar plates. The plate that had no ampicillin in its environment and no plasmid treated E. coli served as a positive control for this experiment because it demonstrated how the E. coli would colonize and grow in a normal setting. The cells in the positive control plate grew into lawn colonies because they were not placed into a selective environment or transformed, so they had no need to acquire ampicillin resistance. Two plates in the experiment contained E. coli cells that were transformed with either the PUC18 or the lux plasmid but were placed in an ampicillin free environment. These two colonies grew

This lab is about moving genes from one thing to another using plasmids. Plasmid has the ability to replicate, so it replicates independently, and separately from the chromosomal DNA. Plasmid are one or more small piece of DNA and they enter cells as a double strand DNA. When they enter the cell as a doubke strand they do not invade he chromosomal DNA. We will also transform bacteria into GFP which is mainly from the jelly fish Aequorea Victoria. The GFP causes the the jelly fish to fluorescent and glow in the dark. After the transformation, bacteria starts to make the GFP which causes them to glow a green color under a ultraviolet light.

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).

70µL of competent E.coli are added to both test tubes; pUC18 and Lux (Alberte et al., 2012). Both test tubes are then tapped and placed back into the ice bath for 15 minutes. While waiting, another test tube is obtained, filled with 35µL of competent cells and labeled NP for no plasmid. A water bath is preheated to 37 degrees Celsius and all three labeled test tubes are inserted into the bath for five minutes (Alberte et al., 2012). Using a sterile pipet 300µL of nutrient broth are inserted into both the control and Lux test tubes and 150µL are inserted to the no plasmid test tube to increase bacterial growth. All three test tubes are then incubated at 37 degrees for 45 minutes. Six agar plates are obtained and labeled to correspond each test tube, three of the plates contain ampicillin. A pipet is used to remove 130µl from each test tube containing a plasmid and insert it into the corresponding agar plate. For this, a cell spreader is first

There were two tubes used in this process: the tube that contained the primary culture and the tube that contained the nutrient agar where the unknown bacteria would grow. First, the inoculating loop was flamed. After removing the caps of both the test tubes, they were flamed to prevent contamination of the unknown bacteria. The inoculating loop was cooled for a few seconds and was then placed into the test tube containing the bacteria. The inoculating loop with the bacteria was placed into the nutrient agar test tube for cultivation. Before the test tubes were capped, they were flamed once again. Also, isolation of the unknown bacteria had to completed. Nutrient agar was placed in the petri dish, and was left to gel for a few minutes. After the agar gelled, the inoculating loop was used to acquire bacteria and streak the unknown onto the plate for

The unknown project was an experiment where the student was given a petri dish of unknown bacteria. Tests were performed on it to identify the specific species. The purpose of the experiment was to learn about the identifying tests, and procedures in the identification of specific microbes. The reason the master plate was used to create a working plate is so that if the working plate becomes contaminated, one can resort back to the master plate for the pure strain of the bacteria and create a new working plate. The purpose of the first procedure, the gram stain, was to be able to dye and then distinguish gram negative and gram-positive cells on a smear. The second procedure, the citrate test is used to see if the bacteria can use citrate as

There of the dishes turned out as expected in this experiment. Our group expected there to be growth in the LB -pGLO dish as the bacteria were not exposed to the antibiotic ampicillin. Furthermore, our group also expected to see inhibited bacterial growth in the LB/amp +pGLO dish as there was ampicillin in the dish, but some of the bacteria were immune as they possessed immunity to the ampicillin. Moreover, our group expected that there would be no bacterial growth in the LB/amp -pGLO dish, as the bacteria were exposed to ampicillin and were not immune. However, the final dish, LB/amp/arbo +pGLO, did not turn out as expected. While it was expected to allow for inhibited bacterial growth and the bacteria to become florescent,

Coli. The first standard E. Coli has no resistance plasmid while the second strain contains a resistance plasmid with genes protecting it from ampicillin. This standard E. Coli and pAMP (plasmid-Ampicillin) E. Coli were each streaked across plates containing the antibiotic and containing growth supportive Lurithea Broth. The purpose of this lab was to test their growth in each medium. Our hypothesis was that while the ampicillin resistant E. Coli would show growth in both LB and LB-AMP plate, the standard E. Coli would only grow in the LB plate for it contains no resistant plasmids against the

While the plate with -pGLO on the LB/AMP agar plate would not have any colonies, and the –pGLO on the LB agar plate would have many colonies of E. coli. The +pGLO on the LB/AMP plate did have two colonies but the only colonies produced on the LB/AMP/ARA plate had a colony not growing on the actual agar plate. The –pGLO on LB/AMP did not have colonies of E. coli growing on it. But the –pGLO on the LB agar plate had many colonies growing on the plate. All of the colonies were clear in color even under the ultraviolet light. Therefore the plate that did not support my hypothesis was the plate with the arabinose sugar, since that was the one thought to grow E. coli do to the ampicillin resistant DNA that was in theory picked up by the E. coli. This plate also should have glowed green under the UV light but instead the colonies remained clear. I hypothesize that E. coli cells recognized the foreign DNA and then destroyed it believing the new DNA would be harmful to the cell. This tendency of bacteria cells to kill foreign DNA could answer be a problem that would explain why we had few colonies that survived in the presence of ampicillin and why none of the colonies in the arabinose glowed green. Also the time the E. coli

At the same time, bacterial colonies are needed to be placed into PCR, and use photograph of gel to determine the size of tetracycline resistance genes to distinguish the bacteria. It is relevant to research 1 because serial dilution is the first step to get colonies' samples that is further being used to distinguish plasmids. It is also relevant to research 2 because serial dilution is also the first step to count the frequency of tetracycline resistant. Week 2: Experiment Part B By placing the bacteria into the PCR machine, much more copies of genes would be made to make them visible under electrophoresis, which can then be compared with control ladder to distinguish the kind of bacteria.

The experiment was a success and the four E Coli plates brought forth the expected results. The two control plates were -pGLo LB and -pGLo LB/amp. On the -pGLo LB plate, there was no ampicillin to prevent the growth of E Coli so the bacteria thrived and grew massively across the plate. There were no specific colonies, rather there was E Coli spread out across the entirety of the gel. This serves as a control because it proves that E Coli was able to grow in the environment provided for it.

Lab Report on pGAL Transformation In order to understand this lab the student first needs to understand how recombinant DNA is formed. To begin, the student extracts a plasmid, which is a circular strand of DNA found within bacterial cells, from the bacteria. Restriction enzymes begin to cut the plasmid at certain sequences of nitrogenous bases.

a. LB+plasmid and LB-plasmid: Both of these plates had a lawn of bacteria. This proves

Plasmids are small double stranded circular non chromosomal DNA molecules containing their own origin of replication. Hence, they are capable of replication independent of the chromosomal DNA in bacteria. Plasmids present in one or more copies per cell, can carry extra chromosomal DNA from one cell to another cell and serve as tools to clone and manipulate genes. Plasmids used exclusively for this purpose are known as vectors. The genes of interest can be inserted into these vector plasmids creating a recombinant plasmid. Recombinant plasmids can play a significant role in gene therapy, DNA vaccination, and drug delivery [Rapley, 2000].

Bacterial transformation is the process of moving genes from a living thing to another with the help of a plasmid.The plasmid is able to help replicate the chromosomes by themselves; laboratories use these to aid in gene multiplication. Bacterial transformation is relevant in everyday lives due to the fact that almost all plasmids carry a bacterial origin of replication and an antibiotic resistance gene(“Addgene: Protocol - How to Do a Bacterial