Premium
An improved method of plant megabase DNA isolation in agarose microbeads suitable for physical mapping and YAC cloning
Author(s) -
Wing Rod A.,
Rastogi Vipin K.,
Zhang HongBin,
Paterson Andrew H.,
Tanksley Steven D.
Publication year - 1993
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1993.04050893.x
Subject(s) - agarose , physical mapping , dna , restriction enzyme , cloning (programming) , biology , isolation (microbiology) , dna extraction , protoplast , molecular cloning , computational biology , microbiology and biotechnology , genetics , gene , gene mapping , bioinformatics , polymerase chain reaction , computer science , chromosome , peptide sequence , programming language
The isolation of high quality megabase DNA from plant cells that is susceptible to a variety of molecular reagents is a critical first step in the physical analysis of complex genomes. A method for the isolation of such DNA by encapsulating plant protoplasts in agarose microbeads is presented. In comparison with the conventional agarose plug method, microbeads provide a dramatic increase in the surface area yielding megabase DNA that can be treated essentially as an aqueous DNA solution. Examples of the utility of DNA prepared by this technique for physical mapping, partial restriction enzyme digestion and cloning of large inserts as YACs are presented.