Research Proposal
Lab Proposal: Lysozyme purification from egg-white.
1. An explanation of why you selected your research topic. Consider issues like:
Q: Why is the topic significant?
A: Every bacteria carries peptidoglycan which is a layer outside the plasma membrane of most bacterias. Peptidoglycan is the specific site that lysozyme targets, lysozyme is an enzyme found in tears, saliva, sweat and other body fluids. It is capable of breaking the chemical bonds in the outer cell wall of the bacteria.
Q: Why is it important to learn more about this topic?
A: Prevent other bacterias from spreading or get rid of a international bacteria.
Q: Why is it important to conduct research in this area?
A: It is important to conduct a research in this area because the advancement of lysozyme can perhaps in the future become more advanced and have a cure to other bacterias.
2. A description of how you will be researching your topic. For example, you might be planning to:
Observe: What I will observe the most is the pallets that are left over when the products are centrifuged. What I will also observe is the lysozyme that was made was pure enough to extract DNA from.
Experiment:
Partial purification of lysozyme
1. For the partial purification of lysozyme, egg whites, carefully separated from the egg yolks, were diluted 3- or 3.3-fold with 0.05 M NaCl solution.
2. To precipitate the egg white proteins other than lysozyme, the pH of this mixture was set to 4.0 by carefully adding several drops of 1 N acetic acid and it was diluted with an equal volume of 40% (v/v) ethanol. After 30 minute incubation at room temperature in the presence of ethanol, the mixture is centrifuged at 15,000 x g for 15 min at 4 °C; then the precipitates were discarded.
3. The supernatants were analyzed for their lysozyme activity and protein content to determine if lysozyme extraction was successful.
4. A DNA extraction is performed using a kit the epicenter QuickExtract Bacterial DNA Extraction Kit (QEB0905T) and E.Z.N.A. Bacterial DNA Kit from Omega bio-tek (D3350-00) as controls
5. A DNA extraction is performed using both kits but replacing the kit provided lysozyme with the student prepared egg-white (EW) lysozyme
3. A Variable Table that includes variable name, type (Independent, dependent, controlled), units, and a description. There is a sample table in this presentation
.
.
Name
|
I/D/C
|
Symbol
|
Units
|
Description
|
Amp (Lysozyme)
|
Dependent
|
_____
|
____
|
Amount of Lysozyme from Egg yolk extraction
|
Lysozyme
|
Independent
|
_____
|
______
|
Method for extraction lysozyme from Chicken Egg
|
Eggs
|
Control
|
______
|
_______
|
Egg from the same day plus the store the eggs were contained from
|
4. An overview of what type of results you anticipate finding. You might include:
Q: Your research question (what you intend to try to support or discover)
A: Is it possible to extract and develop a protocol from lysozyme from egg-yolk?
Q: Your hypothesis
A: I think that the result of purifying Lysozyme will not exactly be pure. And that the Gel Imaging will not come completely pure.
Q:The results that you may discover
A: Lysozyme will come out to be completely pure.
5. At least 5 scholarly sources as references for your project.
References
Sigma- Aldrich. A part of Millipore Sigma. Peptidoglycans. Glycobiology Analysis Manual, 2nd Edition. (2016). Retrieved 16 October 2016
Study.com. Lysozyme: Definition, Function & Structure.(2003-2016). Retrieved 16 October 2016
PDB- 101. Molecular exploration through biology & medicine. Lysozyme. Retrieved 16 October 16 2016.
Xue-li, C., Ting L., & Ito,Y. (2007). Separation of Chicken Egg-White Lysozyme by High- Speed Countercurrent Chromatography using a Reverse Micellar System. Journal of Liquid Chromatography & Related Technologies, 30(17), 25932603
QuickExtract ™ bacterial DNA extraction kit (pdf). (2012, December). Retrieved from
Comments
Post a Comment