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Screening of transgenic plants by multiplex PCR

Marie Mannerlöf, Paul Tenning
Plant molecular biology reporter 1997 v.15 no.1 pp. 38-45
genetic transformation, Zea mays, Helianthus annuus, Petunia hybrida, Capsicum annuum, Cucumis melo, Lactuca sativa, shoots, Solanum tuberosum, transgenic plants, polymerase chain reaction, rapid methods, Cucurbita pepo, Nicotiana tabacum, Brassica napus, Beta vulgaris, Solanum lycopersicum var. lycopersicum
A protocol is described, for the rapid screening of a large number of putative transgenic shoots. Genomic DNA is isolated and screened by PCR. To validate the purity of the DNA, PCR amplification is done with primers homologous to an endogenous gene. Multiplex PCR is used to screen for the transgenic shoots with two sets of primers, one set against the endogenous gene (internal control) and the other set against the gene used in transformation. This protocol has been successfully used on maize, melon, oil-seed rape, pepper, petunia, potato, squash, sugar beet and tobacco.