Abstract
Application of a simplified protocol for nucleic acid preparation for polymerase chain reaction (PCR) detection of greening bacterium (Candidatus Liberibacter asiaticus; Cla) and Citrus mosaic virus (CMBV) associated with citrus is described. Crude extracts of citrus tissues in NaOH - ethylenediaminetetraacetic acid solution, prepared without the use of liquid nitrogen, were spotted on a nitrocellulose membrane (NCM) and then eluted in water at 80 °C for 10 min. Both the greening bacterium and CMBV were detected by PCR from the eluted liquid from the spotted NCMs. The detection efficacy for the templates obtained in the NCM-eluted liquid was comparable with that of templates obtained with a multistep laboratory method or a commercial kit for nucleic acid preparation. The protocol is simple, inexpensive, rapid, and applicable to large-scale surveys of citrus trees. The spotted-membrane methodology can also be used for short-term sample storage for future testing or for long-distance sample transport to a detection laboratory.
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.
