Rapid DNA Extraction Methods for Direct-PCR Detection Citrus Huanglongbing

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[Objective] This study aimed to develop a simple and effective DNA extraction method for citrus Huanglongbing pathogen detection.[Method] From aspects of preparing procedure,prepare time and the quality of DNA,advantages and disadvantages of three sample preparation methods were compared,include two Direct-PCR extraction methods and one universal genomic DNA extraction kit method. In addition,PCR amplification effect on specific primers for 16S rDNA of “ Candidatus Liberibacter asiaticus”( CLas) had also been evaluated.[Result] The results showed that RT-qPCR detected CLas by using DNA obtained from one of Direct-PCR extraction method as templates. Under improved Direct-PCR extraction method,16S rDNA of CLas could also be amplified by routine PCR.[Conclusion]A simple and effective DNA extraction method of citrus Huanglongbing pathogen have been established,which provides technical supports for preparation of large number of samples for detection of CLas. [Objective] This study aimed to develop a simple and effective DNA extraction method for citrus Huanglongbing pathogen detection. [Method] From aspects of preparing procedure, prepare time and the quality of DNA, advantages and disadvantages of three sample preparation methods were compared, include two PCR-PCR methods and one universal genomic DNA extraction kit method. In addition, PCR amplification effect on specific primers for 16S rDNA of “Candidatus Liberibacter asiaticus” (CLas) had also been evaluated. [Result] The results showed that RT-qPCR detected CLas by using DNA obtained from one of the Direct-PCR extraction method as templates. Under improved Direct-PCR extraction method, 16S rDNA of CLas could also be amplified by routine PCR. [Conclusion] A simple and effective DNA extraction method of citrus Huanglongbing pathogen have been established, which provides technical supports for preparation of large number of samples for detection of CLas.
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