Rapid micropreps and minipreps of Ti plasmids and binary vectors fromAgrobacterium tumefaciens View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1995-09

AUTHORS

Xiu-Qing Li, Richard Stahl, Gregory G. Brown

ABSTRACT

A simple and versatile procedure has been developed for the isolation of both large helper/Ti plasmids and binary vectors fromAgrobacterium tumefaciens. Using a slightly modified alkaline lysis protocol, intact plasmid can be recovered from cultures grown in standard micro-centrifuge tubes or culture tubes in sufficient yield and purity to allow for restriction analysis on ethidium bromide stained gels of the >200 kb Ti plasmid DNA. Contamination by chromosomal DNA is minimal and there is thus no need for isopycnic gradient purification. This same procedure can be combined with a high temperature treatment (37°C) and antibiotic selection to generate preparations containing binary vector DNA that are virtually free of interfering Ti plasmid DNA. Restriction patterns produced from these binary vector DNA preparations are unambiguous and therefore preliminary screening by Southern hybridization can be eliminated. More... »

PAGES

349-351

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf01972532

DOI

http://dx.doi.org/10.1007/bf01972532

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1015532966


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