The identification of microalgae based on their morphological features is always challenging due to their similar traits. gaditana strain CCMP527 with a genetic similarity of >99%.
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Molecular identification based on the sequence result and BLAST analysis indicated that direct PCR and manual DNA extraction methods successfully produced high sequences result and confirmed the identity of microalgae species into N. The DNA obtained from both methods were amplified using RbclN primer pair to amplify 1486 bp partial sequence of Nannochloropsis rbcL gene, followed by the sequencing of the PCR product. The same culture was extracted using manual DNA extraction method for comparison. The culture growth was estimated on optical density value and the lysis process was improved using gradual temperature procedure during the PCR process. In this study, Nannochloropsis gaditana NIES-2587 was cultured in f/2 liquid medium.
However, the recent attempts to amplify the rbcL gene of microalga using the previously reported protocol led to poor repeatability. Microalgal direct PCR have been used frequently for species identification, which simplified the DNA isolation procedure. This approach involved preparation of template DNA for polymerase chain reaction (PCR) which is time consuming and requires large amounts of algal cells. Molecular approach plays important role in species identification for microalgae which involves sequencing of specific DNA barcode present in the genome.
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