This revisited Tp conjugation protocol builds on the following two protocols: Conjugation of Thalassiosira pseudonana (DOI: dx.doi.org/10.17504/protocols.io.f55bq86) & Pour plating of Thalassiosira pseudonana (Tp) (DOI: dx.doi.org/10.17504/protocols.io.jfncjme). Reader is strongly encouraged to read both before using this one. Gene-EGFP fusion was expressed under consitutive fcp promoter and terminator.