Batth Rituraj, Cuciurean Ilenuta Simina, Kodiripaka Satish Kumar, Rothman Sarah Skovlund, Greisen Caroline, Simonsen Henrik Toft
Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, 2800 Denmark.
Bio Protoc. 2021 Jan 20;10(19):e3782. doi: 10.21769/BioProtoc.3782.
This protocol describes the generation of protoplasts from protonemal tissue of the moss (syn. ), using Cellulase ONOZUKA R10 and Macerozyme R10, followed by polyethylene glycol (PEG) mediated transformation. The protonemal tissue grown in liquid suspension was harvested and treated with enzyme cocktails mix of 1.5% Cellulase ONOZUKA R10 and 0.5% Macerozyme R10 to generate 1,8 million protoplasts within 3 h.
本方案描述了使用纤维素酶ONOZUKA R10和离析酶R10从苔藓（同义词： ）的原丝体组织中制备原生质体的方法，随后进行聚乙二醇（PEG）介导的转化。收集在液体悬浮液中生长的原丝体组织，并用1.5%纤维素酶ONOZUKA R10和0.5%离析酶R10的酶混合液处理，在3小时内产生180万个原生质体。
