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.