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Screening of transgenic plants by multiplex PCR

Author:
Mannerlöf, Marie, Tenning, Paul
Source:
Plant molecular biology reporter 1997 v.15 no.1 pp. 38-45
ISSN:
0735-9640
Subject:
Beta vulgaris, Lactuca sativa, Zea mays, Cucumis melo, Brassica napus, Capsicum annuum, Petunia hybrida, Solanum tuberosum, Helianthus annuus, Nicotiana tabacum, Solanum lycopersicum var. lycopersicum, Cucurbita pepo, genetic transformation, shoots, transgenic plants, genetic techniques and protocols, polymerase chain reaction, rapid methods
Abstract:
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.
Agid:
1970224