Authors
Chandra K. Singh, Abhishek Ojha, Suchitra Kamle, and Devendra N. Kachru
Summary
The cultivation of transgenic maize (MON810, Monsanto) is widespread globally & exponentially on increase. The international safety regulations mandate use of authentic and stable transgenics and encourage detection methodologies for assessment of the transgenic material, its labeling and rapid identification of spurious GM seed/produce. This study was aimed to develop simpler detection assays with universal assay conditions to characterize the entire transgene cassette in Bt corn (MON810) and verify the insert integrity.
Two distinct PCR assays were developed, employing variable, multiple combinations of primer sets & assay conditions. The PCR assays; one in a standard array & the other in a multiplex format, facilitated concurrent detection of cry1Ab gene sequence (gene specific); maize genome – P-e35S adjoining sequence (event specific); hsp-cry1Ab sequence (construct specific), npt-II marker gene & 02 regulatory (P-e35S promoter & T-nos terminator) sequences- (GMO specific). The standard simplex assay also enabled amplification of maize specific hmgA gene (internal control) and plant specific chloroplast t-RNA gene (positive control). The Limit of detection for cry1Ab gene could be determined simultaneously, by multiplex PCR assay (gene, event and construct specific) and it was 0.1% MON810. A longrun cry1Ab transgene cassette specific PCR assay developed with variable combinations of primer sets, facilitated assessment of structural integrity & stability of the entire transgene cassette of the GM maize. The reported protocol, hence, is an efficient and simple one to provide transgenic maize (MON810) detection under unified assay conditions and elicit all basic information pertaining to GM miaze.
Further details
The protocol was published on Protocol Exchange in 2007. To see the entire protocol, click on the source link.
Advertisement
Stats
- Recommendations n/a n/a positive of 0 vote(s)
- Views 576
- Comments 0