In this project lysozyme
will be extracted from egg whites and compared to commercially produced
lysozyme. Lysozyme is an enzyme that
breaks down glycoproteins (amino acids) on the surfaces of bacteria and kills
them. Large quantities of lysozymes are found in egg whites and can be used in
the process of DNA extraction from bacteria.
The lysozyme extracted from egg whites will be used in DNA extraction to
determine the effectiveness of the lysozyme.
The extracted lysozyme will be utilized in DNA extractions over several
intervals of time after the initial extraction to determine the shelf life of
the enzyme and compared it to commercially produced lysozyme.Research Question:
What is the shelf life of lysozyme extracted from egg whites or will the effectiveness of egg white extracted lysozyme decrease with time in its ability to aid DNA extraction from E. coli?
It is anticipated that the shelf life of egg white lysozyme will produce comparable DNA extraction results from E. coli as the commercially produced lysozyme. Egg white lysozyme will be extracted according to Ganong, (2007). To test the effectiveness, or shelf life, of egg white lysozyme, it will be stored at 4°C for a period of up to two weeks. During these two weeks it will be utilized in a DNA extraction protocol at various intervals from immediately after enzyme extraction to up to two weeks (Fakharany & Hassan, 2016). The effectiveness of the egg white lysozyme to extract DNA will be compared to that of commercially produced lysozyme and it is expected to be comparable in quality and quantity. Quantity and quality of DNA extracts will be determined by DNA electrophoresis and nanodrop spectrophotometry.
Researching the shelf life of egg white lysozyme is important
because egg white lysozyme can substitute expensive commercially produced
lysozyme. Research in DNA extraction can
lead to diagnosis of illnesses and development of new drug treatments. DNA extraction is important in many fields such
as forensics, because the technique can be used generate a full DNA profile.
The efficiency of DNA extraction is crucial in order to perform reliable and
reproducible quantification of the microorganisms in order to identify their
structure and characteristics.
Works Cited
Ganong, B. (2007,
June11). Isolation of Lysozyme from Egg White. Retrieved March 09, 2017, from
Barry Ganong’s Homepage website:
Fakharany, E.E., &
Hassan, M. (2017). A unicersal method
for extraction of genomic DNA from various microorganisms using lysozyme. New Biotechnology, 33(1), S210.
QuickExtractTM bacterial DNA extraction kit
[pdf]. (2012, December). Retrieved March 09, 2017 from:
Verdon, Timothy J., R.
John Mitchell, and Roland A.h. Van Oorschot. "Evaluating the efficiency of
DNA extraction methods from different substrates." Forensic Science
International: Genetics Supplement Series 3.1 (2011): pg 1. Web. 9 Mar.
2017.
Ettenauer, Jörg D.,
Guadalupe Piñar, Ksenija Lopandic, Bernhard Spangl, Günther Ellersdorfer,
Christian Voitl, and Katja Sterflinger. "Microbes on building materials —
Evaluation of DNA extraction protocols as common basis for molecular
analysis." Science of The Total Environment 439 (2012): 44-53. Web.
9 Mar. 2017.
Hey Mary, so during this procedure, are you only going to be working with E. coli instead of your previous identified microbe, Providencia stuartii? To me, it would be kind of disappointing running all those tests on your previous microbe and not being able to do anything else with it. Nonetheless, working with E. coli sounds pretty fun, too!
ReplyDeleteHi Brenda, yes I will only be working with Providencia stuartii from here on out. There was a change of plans in the lab and this is my new experiment microbe friend. Even though I repeated the procedures for identifying the new microbe, it was a lot easier to identify this time around. Identifying the E.coli was good practice and I had more confidence in what I was doing the second time around.
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