If you follow these instructions you will be analyzing mushroom DNA in minutes. I get the sequence to compare by going to http://www.ncbi.nlm.nih.gov/sites/gquery and searching for the species, then clicking nucleotide, then click on a species, this is listed for Psilocybe subaeruginascens: http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=23574923 It says: 1 aaggatcatt attgaataac tttggcgtgg ttgtagctgg ccctctcggg ggcatgtgct 61 cgcctgtcat ctttatatct ccacctgtgc accttttgta gacgtctttg ttggaagctg 121 aataggagag aatgggtgct agtcactctt tctcgagttg aaggctttct caaggtcgct 181 ctatgttttc atatacccca agtatgtaac agaatgtatc tatatggcct tgtgcctata 241 aaactatata caactttcag caacggatct cttggctctc I take out the numbers so I have: aaggatcatt attgaataac tttggcgtgg ttgtagctgg ccctctcggg ggcatgtgct cgcctgtcat ctttatatct ccacctgtgc accttttgta gacgtctttg ttggaagctg aataggagag aatgggtgct agtcactctt tctcgagttg aaggctttct caaggtcgct ctatgttttc atatacccca agtatgtaac agaatgtatc tatatggcct tgtgcctata aaactatata caactttcag caacggatct cttggctctc The paste that into the box labeled "Enter accession number, gi, or FASTA sequence" after going to the main blast.cgi and clicking nucleotide blast. Then go to the Database drop-down and select "Nucleotide collection (nr/nt)" and click BLAST. It gives you a cool graphic that shows the distribution of the blast hits with mouseovers to see the species, and then lists your matches, then shows the actual alignments below. I like to sort the results by total score, and there is a lot of really interesting information on the results page. It says the only 100% match is Psilocybe subaeruginascens, but that Psilocybe cubensis and Panaeolus sphinctrinus are 87% matches! They have DNA from a ton of interesting species from interesting articles that I have never heard of. Psilocybe quebecensis is in there due to an article called "Forensic analysis of hallucinogenic fungi: a DNA-based approach". Psilocybe subaeruginascens in in there from "Phylogenetic relationship of psychoactive fungi based on the rRNA gene for a large subunit and their identification using the TaqMan assay" and from "[Discrimination of psychoactive fungi (commonly called 'magic mushrooms') based on the DNA sequence of the internal transcribed spacer region]". Some of these look like they would be really interesting to try to get ahold of via interlibrary loan. It is unclear how much of a difference is required to define a new species. In my opinion, a consistent 5 base pair difference is enough. Rough instructions for making phylogenetic trees with mafft using only a web browser: 1) Get the DNA from Genbank, http://www.ncbi.nlm.nih.gov/nuccore?term=Panaeolus%20large%20subunit At the bottom right click Send to:, chose file with FASTA format 2) Go to online mafft, start from the FASTA file on your hard drive, open all plots, make sure the lines are red. If any graphs have blue lines remove the sequence from the file, reverse them with rev_comp, then put them back in. http://www.bioinformatics.org/sms/rev_comp.html 3) If sequences do not allign well put the best/longest one at the top 4) Click the phylogenetic tree button Better trees could be made by trimming off unaligned parts and manually verifying that the alignments are sane. Here is a tree I made for Rhodocollybia http://plantobserver.org/rhodocollybia.pdf I made this rough draft phylogenetic tree of Panaeolus using mega5 and information from Genbank. http://plantobserver.org/Panaeolus.phylogenetic.tree.pdf FASTA file: http://plantobserver.org/Panaeolus.fasta From this I may have learned: * Panaeolus papilionaceus and P. sphinctrinus are likely the same species * Panaeolus acuminatus is close to P. olivaceus * Panaeolus cinctulus is close to Panaeolina foenisecii and the split from Panaeolus to Panaeolina is not supported by genetic evidence as Panaeolina does not seem to form a separate clade outside of Panaeolus. * Panaeolus semiovatus is only distantly related to the rest of Panaeolus * Copelandia forms a distinct clade within Panaeolus, and maybe should be a subgenus or section.