Hello phycologists
I was able to isolate about ten isolates of microalgae (cyanobacteria and eukaryotic) from waters polluted by organic matter, the isolation was carried out on BG11 and BBM medium.
The isolates are stored in monoclonal liquid culture, but not axenic, as there is bacterial and fungal contamination.
-my first question: can i go directly to molecular identification without going through morphological identification? "I do not have the expertise for microscopic identification"
-my second question: concerning the molecular identification of microalgae, what are the best primers I can use to identify eukaryotic microalgae and cyanobacteria (could you suggest me some articles).
-My third question: can I run the PCR without having axenic cultures of my isolates, in other words, does the presence of DNA from bacterial and fungal contaminants not influence the PCR results?
Thank you in advance for your answers.
phycological greetings
Depends on what's your purpose. I'd say use ITS as it has the most clearly defined barcode gap between inter- and intraspecific variation.
You dont have to have axenic cultures to perform a pcr. You should use genus specific genes of interest in order to perform pcr for identification (barcoding) your species. In the case of cyanobacteria and algae, use rbcL gene (large subunit of Rubisco).
If you would like to isolate your algae/cyanobacteria, use minimal medium and grow in light. During exponential phase of your algae/cyano, you would be able to assume that these are the main constituents of the consortia, as the other organisms will have difficulties to grow.
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