cloning disaster

I strolled into the lab as eager as one could possibly be this morning. Let me rephrase that: as eager as any scientist could be. I walked directly to the 37 degree centigrade incubator to remove the two E. coli plates that I assumed harbored my recombinant plasmids. I performed several rounds of colony PCR in hopes of amplifying the unique target sequence that I was after, however, this resulted in nothing more than a fluorescent-less agarose gel. These are a pair of N-terminally truncated clones that I need in order to move forward with my research. I guess I'll tell you a little about my research now, since I've gotten the venting out of the way. 

I work on a family of specialized proteins called the Nuclear Factor Y (hereafter referred to as NF-Y). NF-Y are a heterotrimeric transcriptional complex that regulate downstream gene expression. These proteins were originally identified in yeast, and are called Heme Activator Proteins, or simply HAPs, in less advanced eukaryotic systems. These proteins require Iron ions in order to function, similar to other enzymes, namely polymerases, that require magnesium ions. Yeast contain one copy of each of the 3-4 HAP or NF-Y proteins. Conversely, all sequenced higher plants have been shown through bioinformatics to have experienced numerous evolutionary duplication events of NF-Y. For example, Arabidopsis thaliana possess 10 NF-YA, 13 NF-YB, and 13 NF-YC proteins. This evidence suggests that plants have duplicated and undergone slight mutations of these proteins for a broader range of developmental processes.