期刊
VIRUSES-BASEL
卷 11, 期 12, 页码 -出版社
MDPI
DOI: 10.3390/v11121129
关键词
African swine fever virus; ASFV; whole genome sequencing; whole genome amplification; NGS; Illumina; Hungarian ASFV strain; DNAse treatment; whole genome amplification
类别
资金
- National Research, Development and Innovation Office [NKFIH K119381]
- H2020-EU.3.2.1.1. VACDIVA [862874]
- OMA Foundation [101ou6]
- Hungarian Academy of Sciences
In the recent years, African swine fever has become the biggest animal health threat to the swine industry. To facilitate quick genetic analysis of its causative agent, the African swine fever virus (ASFV), we developed a simple and efficient method for next generation sequencing of the viral DNA. Execution of the protocol does not demand complicated virus purification steps, enrichment of the virus by ultracentrifugation or of the viral DNA by ASFV-specific PCRs, and minimizes the use of Sanger sequencing. Efficient DNA-se treatment, monitoring of sample preparation by qPCR, and whole genome amplification are the key elements of the method. Through detailed description of sequencing of the first Hungarian ASFV isolate (ASFV_HU_2018), we specify the sensitive steps and supply key reference numbers to assist reproducibility and to facilitate the successful use of the method for other ASFV researchers.
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