A sequencing protocol of some DNA regions in nuclear, chloroplastic and mitochondrial genomes with an individual colony of Thalassiosira nordenskioeldii Cleve (Bacillariophyceae)

• Main Authors: Iwatani,Naohiro, Murakami,Satoru, Suzuki,Yoshihiro
• Format: Report
• Language: English
• Published: Department of Biological Science, School of Science, Kanagawa University/Department of Biological Science, School of Science, Kanagawa University/Department of Biological Science, School of Science, Kanagawa University 2005
• Subjects: Bacillariophyceae; chelating agent; DNA sequence; micromanipulation; Thallassiosira nordenskioeldii; Polar bioscience
• Online Access: https://nipr.repo.nii.ac.jp/?action=repository_uri&item_id=6223; http://id.nii.ac.jp/1291/00006223/; https://nipr.repo.nii.ac.jp/?action=repository_action_common_download&item_id=6223&item_no=1&attribute_id=18&file_no=1

From cultured cells of micro algal species of Thalassiosira nordenskioeldii Cleve, total DNA was extracted with a buffer containing a chelating agent, Chelex 100. Each DNA fragment for 18s rDNA, 16s rDNA and coxI in nuclear, chloroplastic and mitochondrial genomes, respectively, were amplified from the DNA extracts by polymerase chain reactions, and their sequences were determined. This protocol for determining DNA sequences, was scaled down for the DNA extracts from small amounts of cells. Finally, DNA sequences in genomes were determined with 10 cells collected by a micromanipulation technique. The quantity of cells corresponded to an individual colony of this species. With this protocol, DNA sequences in various regions in genomes of each individual micro algal species living in natural environments can be determined directly, without isolation of strains and cultures.