Problem solving with contamination and data entry

Since my last post, I have completed more PCR and gel electrophoresis with the adult ticks. Everything was going very smoothly until a couple of days ago when I ran a gel and found every single tick came out as positive for the pathogen Ehrlichia chaffeensis. This seemed abnormal and quite frankly, alarming. If all of the tick extracts that were used in that agarose gel were in fact positive, that would have put the prevalence for 2016 above 20% which is much higher than 2017 and 2018. A difference that high did not seem right to me so I knew the odd results must have been due to contamination that occurred along the way. So, I began trying to figure out what to do and after working with my professor we came up with the solution to use new primers. These new primers that I have begun using are longer in base pairs. Therefor, any contamination that could have occurred with the old primers gets discounted as the new primers attach to a longer section of the DNA. I re-ran the contaminated agarose gels and they came out normal again, giving me only 2 positives instead of 32.

Seeing that my summer of research is wrapping up, I am putting a halt on the DNA analysis to finish up data entry and make sure everything is clearly recorded. I have run into a challenge with data entry though. Because the adult ticks from past years were collected by other students, I am running into some discrepancies with the data that was recorded before this summer. Rather than simply adding my data to what already exists, it feels more like I am having to solve a complex puzzle. Certain excel spreadsheets do not contain the same data that others contain, so I am having to go through in order to clarify what is right. While it can be frustrating, I am glad to be having this experience because it is teaching me the importance of consistency in entering data and double checking to make sure everything is correct.

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