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Simultaneous detection of recombinant DNA segments introduced into genetically modified crops with multiplex ligase chain reaction coupled with multiplex polymerase chain reaction.

Authors
Type
Published Article
Journal
Journal of Agricultural and Food Chemistry
1520-5118
Publisher
American Chemical Society
Publication Date
Volume
57
Issue
7
Pages
2640–2646
Identifiers
DOI: 10.1021/jf803361a
PMID: 19265381
Source
Medline
License
Unknown

Abstract

We developed a multiplex polymerase chain reaction (PCR)-multiplex ligase chain reaction (LCR) (MPCR-MLCR) technique as a novel approach for the simultaneous detection of recombinant DNA segments (e.g., promoters, trait genes, and terminators) of genetically modified (GM) crops. With this technique, target DNA regions were amplified by multiplex PCR, the PCR products were then subjected to multiplex LCR as template DNAs, and the LCR products were then analyzed by polyacrylamide gel electrophoresis and subsequent fluorescent scanning. Seven recombinant DNA segments commonly introduced into some GM crop lines were selected as target DNA regions. In addition, another MPCR-MLCR system for the simultaneous detection of three endogenous DNA segments was designed as a positive control test. The specificity and sensitivity of the method were examined. The method allowed us to detect GM crops comprehensively and is expected to be utilized for efficient screening of GM crops into which any one of the seven recombinant DNA segments have been introduced, and for profiling the segments.

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