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Alkali treatment for rapid preparation of plant material for reliable PCR analysis
Author(s) -
Klimyuk Victor I.,
Carroll Bernard J.,
Thomas Colwyn M.,
Jones Jonathan D.G.
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 - 10.1111/j.1365-313x.1993.tb00169.x
Subject(s) - polymerase chain reaction , alkali metal , primer dimer , dna , denaturation (fissile materials) , protocol (science) , chemistry , computational biology , biology , biochemistry , medicine , nuclear chemistry , multiplex polymerase chain reaction , organic chemistry , pathology , gene , alternative medicine
Summary For plant genetics, it would be useful to monitor easily the segregation of different alleles using the polymerase chain reaction (PCR). Preparation of DNA templates from individual plants needs to be rapid and reliable. A one tube protocol is described that involves subjecting plant tissue pieces to alkali, neutralization and heat denaturation prior to PCR analysis, and that proved to be much faster and more reliable than published protocols.