The optimum culture conditions of the local strain Chaetoceros calcitrans were determined to improve biomass and reduce cost of production. Under outdoor culture conditions, higher cell density was attained when the cultures were enriched with Tungkang Marine Research Laboratory (TMRL) medium composed of cheap technical grade reagents and cultured at 25 g L−1 salinity. The cultures were lighted with two 40 W cool-white GE fluorescent tubes (24–35 μmol photon m−2 s−1). Using semi-continuous culture system under established optimum culture conditions, C. calcitrans can be re-cultured thrice and concentrated at each culture cycle using electrolytic flocculation method to produce 4.6 kg m−3 of diatom paste. The viability of concentrated C. calcitrans after 3 months of storage was comparable to live diatom cells. Simple preservation technique by low-temperature storage is convenient for storing algal concentrates for use as starter cultures and for feeding invertebrates. The paste costs USD 8.24 kg−1 inclusive of the assets and flocculation materials for culturing and harvesting the diatom, respectively. This study established the suitable conditions for mass culture of C. calcitrans and produced concentrated diatoms in paste form that is readily available for aquaculture hatcheries at a lower cost.