Protocol:

1. Added 1 microliter of loading dye to ISSR PCR products
2. Loaded onto gel for imaging
3. ran at 60 volts for about an hour and a half

Analysis of Lupinus ITS Sequences

1. Downloaded Lupine_MEC_2017 file from Canvas
2. Opened file in geneious
3. Trimmed then assembled forward and reverse DNA sequences
4. Selected all consensus sequences and made an alignment using default parameters of Muscle.

Results:

Write Up Questions

1. Yes, my tree had one clade with a support value above .85. They were somewhat close together, and they all seemed to be from populations that had yellow flowers.
2. Yes, a few populations that had purple flowers belonged to the same clade as other purple-flowered populations
3. I believe that although ITS sequences can distinguish populations somewhat, the differences in these sequences are not enough to draw solid conclusions phylogenetically. This is because a lot of sequences failed to be grouped into distinct clades with other sequences. As a result of this, it is hard to claim that the ITS is useful enough to group these differing color populations phylogenically.