期刊
JOURNAL OF VIROLOGICAL METHODS
卷 152, 期 1-2, 页码 106-108出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2008.06.003
关键词
large dsDNA viruses; Oryctes rhinoceros nudivirus (OrNV); whole genome amplification; genome sequencing
资金
- Deutsche Forschungs-gemeinschaft [Je245-7]
The genomic sequence analysis of many large dsDNA viruses is hampered by the lack of enough sample materials. Here, we report a whole genome amplification of the Oryctes rhinoceros nudivirus (OrNV) isolate Ma07 starting from as few as about 10 ng of purified viral DNA by application of 4)29 DNA polymerase-and exonuclease-resistant random hexamer-based multiple displacement amplification (MDA) method. About 60 mu g of high molecular weight DNA with fragment sizes of up to 25 kbp was amplified. A genomic DNA clone library was generated using the product DNA. After 8-fold sequencing coverage, the 127,615 bp of OrNV whole genome was sequenced successfully. The results demonstrate that the MDA-based whole genome amplification enables rapid access to genomic information from exiguous virus samples. (C) 2008 Elsevier B.V. All rights reserved.
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