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A rapid and inexpensive method for the direct PCR amplification of DNA from plants
Author(s) -
Bellstedt Dirk U.,
Pirie Michael D.,
Visser J. Christiaan,
Villiers Margaret J.,
Gehrke Berit
Publication year - 2010
Publication title -
american journal of botany
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.218
H-Index - 151
eISSN - 1537-2197
pISSN - 0002-9122
DOI - 10.3732/ajb.1000181
Subject(s) - biology , polymerase chain reaction , dna , applications of pcr , dna extraction , isolation (microbiology) , multiple displacement amplification , recombinase polymerase amplification , computational biology , genetics , loop mediated isothermal amplification , multiplex polymerase chain reaction , microbiology and biotechnology , gene
• Premise of the study : We present a rapid and inexpensive alternative to DNA isolation for polymerase chain reaction (PCR) amplification from plants. • Methods and Results : The method involves direct PCR amplification from material macerated in one buffer, followed by dilution and incubation in a second buffer. We describe the procedure and demonstrate its application for nuclear and plastid DNA amplification across a broad range of vascular plants. • Conclusions : The method is fast, easy to perform, cost‐effective, and consequently ideal for large sample numbers. It represents a considerable simplification of present approaches requiring DNA isolation prior to PCR amplification and will be useful in plant systematics and biotechnology, including applications such as DNA barcoding.