The Attractin Gene

This week has been very prolific, as I accomplished many tasks. I arrived each day to the laboratory with numerous goals in mind and the determination to accomplish those goals. On Wednesday morning alone, I completed three rounds of PCR! Two of my PCR reactions with a new set of primers worked perfectly with the initial set of conditions that I used, so I began the process necessary to sequence those regions of the attractin gene (introns 14 and 27) in the Kenzin population of mice in order to further determine the association of variation within the attractin gene to coat color variation in the rock pocket mice. I had to adjust the conditions of PCR for the other three sets of primers in order to begin sequencing those regions of the attractin gene. For one set of primers, I adjusted the annealing temperature at which the primers bind to the DNA, the Magnesium chloride concentration, and the time at which the reaction is at the annealing temperature; for another set of primers, I ran a new gradient at a different range of temperatures, resulting in a successful amplification of a region of the attractin gene. Another set of primers resulted in nonspecific product that was shorter than the desired product obtained in the reaction; thus, I may have the opportunity to learn the technique of using internal primers that sequence the region within the product of the first PCR reaction. I further continued research on the various components of PCR and the methods of troubleshooting PCR reactions in order to better understand the experimental methods I am currently conducting.

I also began analyzing and cleaning the sequence data from intron 17 that I received last week, which, after an initial examination of the data, seems to reveal an association of variation within the intron to coat color variation in the Carrizozo population of mice; however, I will not be sure that this assumption is accurate until I conduct further examination of all of the sequences from that population. If there is association to coat color, I will then be determining whether the association between variation in attractin and variation in coat color is greater than or less than the association between variation in the agouti gene and variation in coat color in the Carrizozo, NM, population of rock pocket mice.

I have much more to accomplish in the coming weeks in order to complete the tasks above and begin sequencing more regions of the attractin gene, and I look forward to returning to the laboratory next week to continue to make progress in my research.