Cellulase and Macerozyme-PEG-mediated Transformation of Moss Protoplasts

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 cock...

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Bibliographic Details
Published in:	Bio-protocol Vol. 10; no. 19; p. e3782
Main Authors:	Batth, Rituraj, Cuciurean, Ilenuta Simina, Kodiripaka, Satish Kumar, Rothman, Sarah Skovlund, Greisen, Caroline, Simonsen, Henrik Toft
Format:	Journal Article
Language:	English
Published:	United States Bio-protocol LCC 20-01-2021 Bio-Protocol Bio-protocol LLC
Subjects:	Biology Biotechnology Life Sciences Methods Physcomitrium patens (syn. Physcomitrella patens) Protoplastation Cellulase Macerozyme Moss
Online Access:	Get full text
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	2331-8325 2331-8325
DOI:	10.21769/BioProtoc.3782