During this experiment, we can know details about prokaryotic cells. Prokaryotic cell are generally smaller than eukaryotic cell. Prokaryotic cell is present in bacteria. Eukaryotic cells are seen in multicellular organisms. Prokaryotic cells do not have membrane -bound nucleus. Prokaryotic cells do not contain membrane -bound organelles; While Eukaryotic cells contain nucleus and other cell organelles with membrane bound form. The main aim of this experiment is to understand structural and biochemical properties of Prokaryotic cell.
This experiment is divided in three parts. First part is to study bacterial cell walls by using gram staining method. It involves staining method for different types of bacteria. Bacteria are divided in two type based on their cell wall structure. Second part is to learn transferring techniques of bacterial cultures. Transferring of bacterial culture from existing culture to new growth medium is required to maintain cultures that have become overgrowth. Third part is to learn different isolation methods for pure bacterial cultures. Isolation methods are important to study morphology of bacteria such as color, form type such as circular, irregular and spreading, size of bacteria and elevation such as flat or raised.
Hypothesis: If bacteria are gm positive Stephyloccus then it will give purple-blue color with gram stain. If bacteria are gm negative E.coli then it will give no color with gram stain.
Null hypothesis: Both types of bacteria give same color with gram stain.
Independent variable: bacterial cell wall structure
Dependent variable: color of bacteria
Standard variable: reagent concentration, bacterial culture growth, sterile condition
Materials: agar slants, Petri dish, staining dye, alcohol, safranin, slides, dropper, inoculating loop, spreading rode, different bacterial culture broth, microscope, and incubator.
Method for gram staining: Two clean glass slides were labeled as "E" and "S". Slides were handled by using tissue paper. A wire inoculating loop was sterilized in hot flame. By using sterilized loop two small drops of tap water was put on two slides. Then loop was sterilized again. After proper cool, a small amount of growth was removed from "E.coli" agar-slant culture by using sterilized inoculating loop. A neck of agar-slant was sterilized in flame before closing and after opening. "E.coli" culture which was on loop was transferred on slide "E". The drop of culture was mixed properly with drops of water on slide. It was milky appearance. It was allowed to air dry. After that by using forceps slide "E" was fixed in burner flame by using heat fix method. Above procedure was repeated for slide "S". Only difference was instead of "Escherichia .coli" culture for Slide "S" a culture of...