Abstract
A loop-mediated isothermal amplification (LAMP) and real-time LAMP based assays were developed for quick and sensitive detection of transgenic black pepper plants. Primers (six each) were designed based on the nucleotide sequence of two target regions [kanamycin and Cauliflower mosaic virus (CaMV) 35S promoter] integrated into the genome of transgenic black pepper. Both assays successfully detected the transgenic plants and no cross-reaction was recorded with non-transgenic plants. The sensitivity of LAMP was up to 104 times that of conventional PCR while real-time LAMP was up to 103 times that of LAMP and 107 times to that of PCR. The addition of 6 mM magnesium sulphate and 0.4 M betaine with 1 h reaction time proved optimal for amplification through LAMP assay. The assays were validated by testing putative transformants of black pepper. The present study clearly established that LAMP and real-time LAMP assays can provide a rapid and simple approach for screening transgenic black pepper and other plants transformed by using the above target gene sequences.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
