Creating Poster for Presentation

This week’s lab hours were dedicated to completing my presentation poster. This is my first research poster and it will also be my first time presenting.  I’m looking forward to coming full circle in this S-Stem program, complete my research project and discuss with others what I’ve learned in the process.  Below are portions of the poster completed. 

Summary of Results

Data collected was inconsistent, the following observations were made during the experiment process that could account for the inconsistencies in the data.  In centrifuge tube 4/7/17 Egg White 2 a gnat was found trapped inside contaminating the sample.  Centrifuge tubes were not all flicked/tapped to mix contents evenly before adding samples on the Nano Drop Machine and obtaining data.  The final volume in each centrifuge tube was not consistent due to different size DNA cell pellets formed when centrifuged. This is possibly due to insufficient incubation time of Providencia stuartii TSB, used in the extraction of DNA.  The final observation made was of the DNA yield numbers in the commercial kit samples, they were lower than expected.  This was due to the low concentration of the Ready-Lyse-Lysozyme being used, to correct this the concentration was increased.    

Conclusion

Providencia stuartii DNA yields extracted using egg white lysozyme was detected in samples collected the day of and samples that were 1, 6, and 8 days old.  

The method used in this experiment of using a Nano Drop machine yielded data that was too inconsistent to make a definitive conclusion. 

The results of this study indicated the importance of technique application and that of using more than one method to analyze data in order to have consistent and accountable data.

Further testing using different methods such as PCR and/or gel electrophoresis should be done to further analyze the data and make a definitive conclusion.

Implemented Adjustments to Protocol

The eggs used in this week’s protocols had an expiration date of 04/01/17.  Per my mentor, to check if the eggs were still usable a simple test was performed.  A raw egg was gently dropped into a beaker of water to see if it would float (it didn’t float).  If it did float it meant the egg was no longer usable and if it stayed at the bottom of the beaker it was usable. Also, another egg from the same carton was boiled and gently dropped in a beaker full of water and it did not float. The eggs used were carefully examined when cracked open to check for any discoloration or bad smells both eggs used in the protocols appeared usable.  Due to last week’s observations the Ready-Lyse-Lysozyme used in the commercial kit was increased to a higher concentration to adjust the DNA yields.  This was done because the DNA yields from the commercial kit were lower than expected.  After completing the protocols from start to finish and running the samples (Egg White, Kit, and Water Lysozyme) in the Nano drop machine the DNA yield numbers from the commercial kit increased. Also each centrifuge tube was flicked/tapped to mix contents, the volume of each centrifuge was relatively equal and the Providencia stuartii TSB incubation period was increased to 48 hours.  Next week the samples collected will be put through gel electrophoresis and polymerase chain reaction.  I’m excited to learn some new procedures and hopefully the data collected from these tests will be more consistent.

Analyzing Data Collected From Protocols and Accounting for Inconsistencies

Egg White 3 Gnat

This week in preparing for the rough draft research paper due, the protocols were performed as normal, from start to finish.  The samples (Egg Whites, Kit, Water) were ran in the Nano drop machine, then all the data from the previous weeks tests were collected and analyzed.  With my mentor Josh next to me, watching as I reran all of my samples, observations were made that might account for the inconsistencies in my data.  For example in egg white 3 centrifuge tube on 4/7/17, a gnat was found trapped inside the centrifuge tube, which might account for the abnormal reading in that particular tube. Also before each sample is drawn into a pipette and added onto the Nano drop machine, each centrifuge tube needed to be flicked/tapped so as to mix the contents well and get an accurate, consistent reading.  This is something I wasn’t doing to every centrifuge tube.  Another observation made was that not all centrifuge tubes had the same volume, some had more than others.  The reason for this was because when the samples were centrifuged, the DNA pellets formed were bigger in some and smaller in others.  When the distilled water supernatant was removed from each centrifuge tube, I was careful not to disturb the big or small DNA pellets and left different amounts of distilled water in each tube. From here on out I will work towards having the same volume in each centrifuge tube.  A possible explanation for the different sizes of DNA pellets, might be due to the amount of time the microbe Providencia stuartii was allowed to incubate and grow before being used.  If the TSB tube inoculated with Providencia stuartii was allowed to incubate for only 24 hours the DNA pellet formed was smaller.  Once the incubation time was increased to 48 hours the DNA pellet size increased.  From here on out the incubation period will be 48 hours.  The last observations made were the DNA yield numbers in the commercial kit samples, they were all lower than expected.  To correct this, the concentration of the Ready-Lyse-Lysozyme used in the commercial kit was increased.  Next week when all of these adjustments are implemented and the experiment is run from start to finish, my goal is to have a little more consistency in DNA yields from my samples.