Bacterial Transformation
1. a. –pGLO LB plate
It is expected there to be a lawn growth on this plate and it will not fluorescent under UV light (1). b. –pGLO LB/amp plate It is expected there to be no growth on this plate and it will not fluorescent under UV light (1). c. +pGLO LB/amp plate It is expected there to be colonies growth and it will not fluorescent under UV light (1). d. +pGLO LB/amp/ara plate It is expected there to be colonies growth and it will fluorescent under UV light on this plate (1). e. +pGLO LB/amp liquid culture It is expected there to be growth with turbid appearance and it will not fluorescent under UV light (1). f. +pGLO LB/amp/ara liquid culture
It is expected there to be growth with turbid appearance and it will fluorescent under UV light (1). 2.
Plates/Liquid Culture Type of Growth Fluorescent under UV light?
–pGLO LB plate Lawn growth No
–pGLO LB/amp plate No growth No
+pGLO LB/amp plate Colonies growth; satellite colonies around the mother colony No
+pGLO LB/amp/ara plate Colonies growth; satellite colonies around the mother colony Yes
+pGLO LB/amp liquid culture Cloudy growth and pastel yellow in color No
+pGLO LB/amp/ara liquid culture Cloudy growth and pastel yellow in color Yes
Table 1 – Growth Results on Plates and Liquid Culture 3.a. For the –pGLO LB plate, there was a lawn growth as expected. Although the plasmid pGLO is absent, Escherichia coli bacteria is still present to grow on the
Test tube six contained an elodea and was placed in the dark. It was hypothesized that, after a day, the test tube would be yellow in color. Upon observing, the hypothesis was proven correct because test tube six had remained yellow. This is because the elodea was placed in the dark where it could not receive light to photosynthesize and remove carbon dioxide from the
Two treatment groups were covered with a screen in order to reduce light intensity. Both groups were kept under a controlled light source for fourteen days and plant counts were taken at regular intervals.
7. We added 1.0 mol dm-3 of NaOH into tube #8 and than added 1.0 mold dm-3 of mold dm-3 HCl until a change in color is observed.
To perform this test, a small drop of water is placed on a clean microscope slide. A metal loop that has been properly sterilized in the blue flame and allowed time to cool is used to
At the start of this experiment we were required to obtain a set of four Wisconsin Fast Plants, which are genetically, known as Brassica rapa. These plants have been, “originally selected under continuous fluorescent light to grow and reproduce quickly for research purposes, these petite, fast-growing plants have been used for teaching biology concepts” (Wisconsin Fast Plants). These four pots that contain our plants will be under our watch for the next 16 weeks where we will show our results at the end of the semester.
22. + DNA/LB/AMP plate looked white when the 2nd plates color showed green with a UV light and white in room light.
14. Use the same loop and technique to spread the same cell suspension (+) on the LB+AMP agar plates. Dispose of the sterile loop in a beaker of germicide.
To start off unknown #3, I picked out O17 from rack A and wrote down my name, date, class, and I first noticed the broth was a light yellow that was quite cloudy or milky. Soon later I streaked my organism onto a TSA plate and incubated it at 37oC for 24 hours, t was almost a perfect plate. There was no contamination; I could tell it there was only one species on the plate and that it was okay for me to move on. There were many isolated colonies in quadrants 3 and 4. The colonies were small and white in color, it was not translucent. I then began to make my 2 slants, inoculating a straight line on each slant, I incubated it at 37oC for 24 hours. The next day, I made sure my slants were pure and also there was a thick line of organism where I had inoculated with a yellow-white color. Now I was ready to begin making my slides and start staining. I used E. Coli and S. Aureus next to my unknown to help me
5.) Given that white light contains all colors of the spectrum, what growth results would you expect under white light?
You can grow microorganisms in liquid media or on solid media. Most bacteria can be grown in labs as long as the media contain a source of the major nutrients; carbon, nitrogen, sulphur, and phosphorous. They may also need to have other nutrients. These nutrients are made into a broth and the pH and salinity can be adjusted. The nutrient broth is placed in test tubes, which are plugged with cotton wool, capped with foil and then sterilised in an autoclave, at 121c for 20minutes. These tubes are then cooled before they are inoculated. To prepare the streak plates we dipped an inoculating loop into ethanol then placed it in the flame until the loop glowed red. Still holding the inoculating loop by its handle we removed the lid from
For day three of the unknowns project I observed the growth on all four TSA slants and the difference in growth between colony 1 and 2. On the TSA slants there was a clear distinguish in the growth between colony 1 and colony 2. Colony 1 had a white/yellow layer of growth. Colony 2 had a clear/white color of growth on the TSA slant. There was no pigment that was produced from the colony 2 organism, so there was a negative result.
Purpose: The purpose of this experiment is to gain knowledge of the functions and operations of the compound light microscope and an immersion oil lens by observing prepared slides of various bacteria and blood slides. We are also learning to indentify and observe the various shapes and characteristics of bacteria, as well as, yogurt cultures (fresh and prepared) and blood samples under a microscopic view. We will also be able to distinguish between blood cultures and bacteria specimens.
In the 10-3 pasteurized sample, the plate exhibited 71,000 cells/mL. The results of the additional dilution samples contained too few colony forming units to count. However, in the 10-7 dilution, although the plate demonstrated 12 colonies, there should have been no colony forming units on this plate. The reasons for this could have been that this sample was contaminated from “double-dipping” the sample before dispensing it onto the plate or when using the pipette, it mistakenly was inserted in a higher concentration sample and then immediately to a lower concentration sample before it was dispensed onto the plate.
Preparing serial dilutions of the alfalfa water will ensure an accurate count. The plates are incubated until you see visible colonies, usually 18-24 hours. The colonies you see growing on the plate are considered to have started from one viable bacterial unit but because bacteria are usually not found as individuals, the colony you see may have started from a single cell or a group of cells. The results are reported as colony forming units (CFU’s).
When viewing the unstained cheek cells, the amount of light did not have an effect on viewing the slide; meaning that the cells were not visible until they were stained.