Genomic DNA was isolated from 50 ml of a two-week-old algal cell culture using the Gene Elute™ Plant Genomic DNA Miniprep Kit (Sigma–Aldrich, Hamburg, Germany) according to the manufacturer`s instructions. The chloroplast-encoded gene (rbcL) was amplified from genomic DNA using the proof reading polymerase Phusion (Finnzymes, Thermo Scientific, Schwerte, Germany) according to the manual. The primers used for amplification and the protocol of amplification are the same as in Witkowski et al.56 (link) specified for rbcL sequence.
Witkowski A., Gomes A., Mann D.G., Trobajo R., Li C., Barka F., Gusev E., Dąbek P., Grzonka J., Kurzydłowski K.J., Zgłobicka I., Harrison M, & Boski T. (2015). Simonsenia aveniformis sp. nov. (Bacillariophyceae), molecular phylogeny and systematics of the genus, and a new type of canal raphe system. Scientific Reports, 5, 17115.
Other organizations :
University of Szczecin, University of Algarve, Royal Botanic Garden Edinburgh, Institute for Research and Technology in Food and Agriculture, Institute of Biology of Inland Waters named Ivan Dmitrievich Papanin, Goethe University Frankfurt, Warsaw University of Technology, University of Nebraska–Lincoln
Genomic DNA isolation method (using the Gene Elute™ Plant Genomic DNA Miniprep Kit)
dependent variables
Amplification of the chloroplast-encoded gene (rbcL) from genomic DNA using the Phusion polymerase
control variables
Algal cell culture age (two-week-old)
Volume of algal cell culture (50 ml)
Amplification primers and protocol (same as Witkowski et al.)
controls
No positive or negative controls were explicitly mentioned in the input protocol.
Annotations
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