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Screening of transgenic plants by multiplex PCR

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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.

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Abbreviations

CTAB:

hexadecyltrimethylammonium bromide

nad5 :

gene encoding NADH dehydrogenase

PCR:

polymerase chain reaction

uidA :

β-glucuronidase

surA/surB :

acetolactate synthase

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Authors and Affiliations

  1. Department of Biotechnology, Box 302, S-261 23, Landskrona, Sweden

    Marie Mannerlöf & Paul Tenning

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  1. Marie Mannerlöf
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  2. Paul Tenning
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Correspondence to Marie Mannerlöf.

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Mannerlöf, M., Tenning, P. Screening of transgenic plants by multiplex PCR. Plant Mol Biol Rep 15, 38–45 (1997). https://doi.org/10.1007/BF02772111

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  • DOI: https://doi.org/10.1007/BF02772111

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