Comparative genomics reveals signature regions used to develop a robust and sensitive multiplex TaqMan real-time qPCR assay to detect the genus Dickeya and Dickeya dianthicola

Shefali Dobhal; Gamze Boluk; Brooke Babler; Michael J. Stulberg; John Rascoe; Mark Nakhla; Toni A. Chapman; Alex B. Crockford; Michael Melzer; Anne M. Alvarez; Mohammad Arif

The capability of the developed assay was tested with naturally infected potato samples collected from the field. A total of twenty-four plant samples were brought to the lab and washed with sterile water; stem explant of 1 cm size was taken in an Eppendorf tube and crushed with small pestle; DNA was isolated using the Wizard Genomic DNA purification kit. The DNA was quantified using Nanodrop and used for the multiplex TaqMan real-time qPCR. The multiplex TaqMan qPCR was performed using the reaction components and conditions as described above. Three replicates of each sample were run along with the positive and the negative controls.